Patent application title: Human Salty Taste Receptor And Methods Of Modulating Salty Taste Perception
Inventors:
Joseph G. Brand (Wayne, PA, US)
Joseph G. Brand (Wayne, PA, US)
Taufiqul Huque (Philadelphia, PA, US)
Assignees:
MONELL CHEMICAL SENSES CENTER
IPC8 Class: AG01N33566FI
USPC Class:
436501
Class name: Chemistry: analytical and immunological testing biospecific ligand binding assay
Publication date: 2014-04-03
Patent application number: 20140093978
Abstract:
Methods for identifying modulators of the epithelial sodium ion channel
and for identifying modulators of salty taste perception are described.
Also featured are isolated human salty taste receptors, artificial lipid
bilayers comprising an epithelial sodium ion channels, and kits for
practicing the claimed methods.Claims:
1. A method for identifying modulators of epithelial sodium ion channels,
comprising: assembling at least one epithelial sodium ion channel in an
artificial lipid membrane, wherein the epithelial sodium ion channel
comprises at least three subunits, wherein the subunits are independently
an alpha subunit, a beta subunit, a gamma subunit, a delta subunit or an
epsilon subunit; contacting the ion channel with a test compound in the
presence of sodium or lithium; and determining a modulation of the
biological activity of the epithelial sodium ion channel in the presence
of the test compound relative to the biological activity of the
epithelial sodium ion channel in the absence of the test compound.
2. The method of claim 1 wherein the epithelial ion channel comprises at least one delta subunit, at least one beta subunit, and at least one gamma subunit.
3. The method of claim 1, wherein the epithelial ion channel comprises at least one alpha subunit, at least one beta subunit, at least one delta, and at least one gamma subunit.
4. The method of claim 3, further comprising at least one epsilon subunit.
5. The method of claim 1, further comprising contacting the epithelial sodium ion channel with an epithelial sodium ion channel antagonist.
6. The method of claim 1, wherein the artificial lipid membrane is a micelle, liposome, or lipid bilayer.
7. The method of claim 1, wherein at least two subunits of an epithelial sodium ion channel are present in the lipid membrane at differing ratios relative to each other.
8. The method of claim 1, wherein the epithelial sodium ion channel comprises at least one biological activity of a functional human salty taste receptor.
9. A method according to claim 1, adapted for high throughput screening.
10. A planar lipid bilayer comprising at least one type of phospholipid and an epithelial sodium ion channel or specific ratios of epithelial sodium ion channel subunits, wherein the subunits are independently an alpha subunit, a beta subunit, a gamma subunit, a delta subunit, or an epsilon subunit.
11. The artificial lipid membrane of claim 10, wherein the membrane is a micelle, liposome, or lipid bilayer.
12. The planar lipid bilayer of claim 10, comprising at least one human epithelial sodium ion channel comprising at least one alpha subunit, at least one beta subunit, at least one delta subunit, and at least one gamma subunit.
13. The planar lipid bilayer of claim 12, further comprising at least one epsilon subunit.
14. The planar lipid bilayer of claim 10, comprising at least one epithelial sodium channel comprising at least one delta subunit, at least one beta subunit, and at least one gamma subunit.
15. The planar lipid bilayer of claim 14, wherein the at least one epithelial sodium channel further comprises at least one alpha subunit.
16. The planar lipid bilayer of claim 10, wherein the epithelial sodium channel comprises at least one delta subunit.
17. The planar lipid bilayer of claim 16, wherein the epithelial sodium channel further comprises at least one alpha subunit.
18. The planar lipid bilayer of claim 10, wherein the epithelial sodium channel comprises at least one delta subunit, at least one beta subunit, and at least one gamma subunit wherein the ratio of the delta, beta, and gamma subunits to each other is 1:1:1.
19. The planar lipid bilayer of claim 18, wherein the epithelial sodium channel further comprises at least one alpha subunit.
20. A method for preparing the artificial lipid membrane of claim 10, comprising: admixing a micelle or liposome comprising at least one phospholipid with an epithelial sodium ion channel or specific ratios of epithelial sodium ion channel subunits, wherein the epithelial sodium ion channel or epithelial sodium ion channel subunits are dissolved in a suitable aqueous buffer comprising at least one surfactant; incubating said micelle or liposome with the epithelial sodium ion channel or epithelial sodium ion channel subunit for a sufficient amount of time; and removing the at least one surfactant.
21. The method of claim 20, further comprising reconstituting the proteo-liposome into a planar lipid bilayer.
22. A method for identifying modulators of salty taste perception, comprising: assembling at least one epithelial sodium ion channel in an artificial lipid membrane, wherein the epithelial sodium ion channel comprises at least one beta subunit, at least one gamma subunit, and at least one delta subunit; contacting the ion channel with a test compound in the presence of sodium or lithium; determining a modulation of the biological activity of the epithelial sodium ion channel in the presence of the test compound relative to the biological activity of the epithelial sodium ion channel in the absence of the test compound; and administering the test compound to a subject and determining a modulation of salty taste perception in the subject relative to the level of salty taste perception in the subject in the absence of the test compound.
23. The method of claim 22 wherein the delta subunit comprises the amino acid sequence of SEQ ID NO:12.
24. The method of claim 22 further comprising the step of screening positive candidate compounds in a cell-based assay for epithelial sodium channel activity.
25. An isolated human salty taste receptor comprising at least one beta polypeptide subunit, at least one gamma polypeptide subunit and at least one delta polypeptide subunit, wherein said delta polypeptide subunit comprises the amino acid sequence of SEQ ID NO:12.
26. The isolated human salty taste receptor of claim 25 wherein said delta polypeptide subunit has the amino acid sequence of SEQ ID NO:9.
27. A kit for identifying modulators of the human salty taste receptor comprising: at least one phospholipid; substantially purified epithelial sodium ion channel subunits comprising delta subunits, beta subunits, and gamma subunits; optionally comprising an epithelial sodium ion channel modulator, sodium or lithium; and instructions for using the kit in a method for identifying modulators of the human salty taste receptor.
28. The kit of claim 27 wherein the subunits are admixed in known ratios in a single container.
29. The kit of claim 27 wherein at least two subunits are present at differing ratios relative to each other.
30. The kit of claim 27, wherein the modulator is amiloride, phenamil, benzamil, chlorhexidine or a source of guanidinium ion or an organic polyamine.
31. A compound identified by the method of claim 1, wherein the compound activates the epithelial sodium ion channel and induces a salty taste perception when administered to the mouth of a subject, provided that the compound is not sodium or lithium.
Description:
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is continuation of U.S. application Ser. No. 13/008,330, filed Jan. 18, 2011, which is a divisional of U.S. application Ser. No. 11/875,200, filed Oct. 19, 2007, which claims benefit to U.S. Provisional Application No. 60/853,290 filed Oct. 19, 2006, the entire contents of which are incorporated by reference herein, in their entirety and for all purposes.
FIELD
[0002] The invention relates generally to the field of cell biology. More specifically, the invention relates to sodium ion channels and their role in the recognition of salty taste in humans.
BACKGROUND
[0003] Various publications, including patents, published applications, technical articles and scholarly articles are cited throughout the specification. Each of these cited publications is incorporated by reference herein, in its entirety and for all purposes.
[0004] Sodium plays an important role in the body's metabolism, including, among other things, electrical impulse transmission and fluid and electrolyte homeostasis. In addition, sodium contributes to the development and stability of flavor in the various foods ingested by animals, particularly by humans. The sodium ion can inhibit the bitter taste of some stimuli, thereby modifying the taste of food. This inhibitory effect of sodium on bitter taste does not depend on the saltiness of the compound containing the sodium ion, but rather depends on the concentration of the sodium ion.
[0005] Excess intake of sodium, however, has been implicated in various disease states, including gastric cancer and hypertension. Hypertension is a major risk factor for heart disease, stroke, and kidney disease. Because of the potential negative health effects of excess sodium consumption, the United States FDA recommends that adults limit their intake to less than 2400 milligrams of sodium per day. Nevertheless, Americans generally far exceed this recommended allowance. As such, various medical and scientific groups have recommended drastic reductions in sodium intake.
[0006] To further the goal of reduced sodium intake, numerous salty taste mimics and salty taste enhancers have been developed. In general, such mimics have not proven commercially viable as they lack the clean saltiness of sodium chloride, and most do not affect food flavor as sodium salt does.
[0007] The dearth of mimics of salty taste, commonly known as salt substitutes, reflects the extreme structural specificity of the taste receptor. As far is known, only sodium chloride (NaCl) and lithium chloride (LiCl) impart a true salty taste. Both heavier anions paired with Na and Li, and heavier cations paired with Cl tend to be bitter. The cation specificity suggests an ion channel, while the chloride effects suggests paracellular shunts. In addition, the concentration at which NaCl imparts a salty taste is above 50 mM, a concentration on the higher end of receptor processes. These two observations--the specificity for Na and Li, and the effective concentration range--are believed to be the key to discovering the mechanism of salty taste in humans.
[0008] Over the past two decades, numerous studies, both qualitative and quantitative, of salt-induced changes in neural activity in the presence or absence of specific inhibitors and enhancers have led to the supposition that an epithelial sodium channel (ENaC) acts as the primary receptor for saltiness (Brand et al. (1985) Brain Res. 334:207-14; Feigin et al. (1994) Am. J. Physiol. 266(Cell Physiol):C1165-72; and, Brelin et al. (2006) Adv. Otorhinolaryngol. 63:152-90). While the ENaC serves as the salt receptor for many experimental animals (Halpern, B P (1998) Neurosci. Biobehav. Rev. 23(1):5-47), no conclusive evidence has emerged that the same holds true for human beings. Notably, the inability of amiloride to inhibit sodium-induced salty taste response in humans suggests that ENaCs are not involved in human salty taste recognition, at least to the extent observed in other animals.
[0009] Because of this discrepancy between human and animal models, the transduction mechanisms underlying the perception of salty taste in humans remain under investigation. Sufficient activation of the nerve eventually evokes the sensation of saltiness in the higher cortical areas (Schoenfeld, Mass. et al. (2004) Neuroscience. 127:347-53).
[0010] Because of the robust response shown to amiloride by taste cells of many rodents, the ENaCs in these cells are assumed to be located primarily at the apical membrane, above the level of the tight junctions. This location makes them susceptible to the action of drugs such as amiloride. It is assumed that amiloride cannot pass the tight junctions. Augmenting the direct mechanism at the apical membrane is a paracellular shunt pathway into the basolateral area of taste buds below the tight junction level (Mierson, S et al. (1996) J. Neurophysiol. 76:1297-309). Since sodium can pass the tight junctions, the paracellular mechanism should result in an amiloride insensitive salty taste response. The human salty response may be amiloride-insensitive because the vast majority of taste cell ENaCs are located below these tight junctions. Other mechanisms for salt perception may exist. These could be entirely different from the ENaC, or an alternative manifestation of the ENaC due to sodium load or hormonal influences on ENaC expression or composition.
[0011] ENaCs comprise a family of cation channel proteins mediating sodium permeation in epithelia (Mano, I et al. (1999) Bioessays 21:568-78). Expression cloning originally demonstrated that there are three homologous genes, each encoding one of the three subunits of the channel--i.e., alpha (α), beta (β) and gamma (γ) (Canessa, C M et al. (1994) Nature 367:463-7). Co-expression of all three subunits is essential for maximal Na+ channel activity, although the alpha subunit by itself produces a small current. A fourth subunit, delta (δ) was later cloned and shown to be similar to the alpha subunit both structurally and functionally, albeit with a 30-fold lower affinity for amiloride (Waldmann et al. (1995) J. Biol. Chem. 270:27411-4). This lower amiloride sensitivity is assumed to be reflected in a motif called the PreMR2 sequence. The transmembrane topology of the ENaCs comprises two hydrophobic transmembrane domains flanking a long extracellular loop, with intracellular amino and carboxyl termini. The subunit stoichiometry of the ENaCs may be species-specific and tissue-specific, since there is evidence for an α2βγ configuration in rats (Firsov et al. (1998) EMBO J. 17:344-52) and an (α)1β(1)γ(1) arrangement in humans (Staruschenko, A (2005) Biophys. J. 88:3966-75).
[0012] For improved health and wellness, there is a need to diminish sodium intake. This need must be balanced with the desire for the taste of sodium, and the ability of sodium to impart improved flavor in food. One attractive means to diminish dietary sodium without sacrificing sodium flavor is to use modulators of salty taste. Thus, there is a need to establish the definitive receptor for salty taste perception and for a means to identify modulators of salty taste perception.
SUMMARY
[0013] The invention provides an isolated human salty taste receptor comprising at least one beta polypeptide subunit, at least one gamma polypeptide subunit, and at least one delta polypeptide subunit wherein said delta polypeptide subunit comprises the amino acid sequence of SEQ ID NO:12. In some aspects, the delta polypeptide subunit has the amino acid sequence of SEQ ID NO:9. Also provided is an isolated human salty taste receptors comprising at least one alpha polypeptide subunit, at least one beta polypeptide subunit, at least one delta polypeptide subunit, and at least one gamma polypeptide subunit.
[0014] The invention also provides a method for identifying modulators of epithelial sodium ion channels. Such methods include assembling at least one epithelial sodium ion channel in a lipid membrane (wherein the epithelial sodium ion channel comprises at least three types of subunits, which are independently an alpha subunit, a beta subunit, a gamma subunit, a delta subunit, and an epsilon subunit); contacting the ion channel with a test compound in the presence of sodium ions or lithium ions; and determining a modulation of the biological activity of the epithelial sodium ion channel in the presence of the test compound relative to the biological activity of the epithelial sodium ion channel in the absence of the test compound. The lipid membrane is preferably an artificial membrane.
[0015] In some aspects, the epithelial ion channel comprises one alpha subunit, one beta subunit, and one gamma subunit. In other aspects, the epithelial ion channel comprises one alpha subunit, one beta subunit, one gamma subunit, and one epsilon subunit. In other aspects, the epithelial ion channel comprises two alpha subunits, one beta subunit, and one gamma subunit. In further aspects the epithelial ion channel comprises three alpha subunits, three beta subunits, and three gamma subunits. Additional aspects include those wherein the epithelial ion channel comprises one delta subunit, one beta subunit, and one gamma subunit. In other aspects, the epithelial ion channel comprises two delta subunits, one beta subunit, and one gamma subunit. In still further aspects, the epithelial ion channel comprises two delta subunits, two beta subunits, and two gamma subunits. In still further aspects, the epithelial ion channel comprises three delta subunits, three beta subunits, and three gamma subunits.
[0016] In the method for identifying modulators of epithelial sodium channels, the method may further include contacting the epithelial sodium ion channel with an epithelial sodium ion channel antagonist, such as, but not limited to chlorhexidine, amiloride, phenamil, benzamil or a homolog, analog, or derivative thereof.
[0017] In the method for identifying modulators of epithelial sodium channels, suitable lipid components for the membrane include at least one of phosphatidylcholine, phoshpatidylethanolamine, phostphatidylserine, phosphatidylglyine, phosphatidylinositol, sphingomyelin, cholesterol, cardiolipin, or a homolog, analog, or derivative thereof. As such the lipids may be organized as a micelle, liposome, or lipid bilayer.
[0018] In some aspects of the method for identifying modulators of epithelial sodium channels, at least two subunits of an epithelial sodium ion channel are present in the lipid membrane at differing ratios relative to each other.
[0019] In the step for determining a modulation of the biological activity of the epithelial sodium ion channel, any suitable means known in the art may be used, such as, but not limited to, voltage clamping, and/or measurement of an indicator dye. The method may be adapted for high throughput screening.
[0020] The method for identifying modulators of epithelial sodium channels thus provides compounds identified by the method that act as modulators of the epithelial sodium channels. These compounds may be formulated into compositions by admixing the compounds with a pharmaceutically acceptable carrier.
[0021] In a specific aspect, the invention provides a method for identifying modulators of the human salty taste receptor comprising: assembling at least one salty taste receptor in a lipid membrane, wherein the salty taste receptor comprises at least one beta subunit, at least one gamma subunit, and at least one delta subunit; contacting the ion channel with a test compound in the presence of sodium ions or lithium ions; and determining a modulation of the biological activity of the salty taste receptor in the presence of the test compound relative to the biological activity of the salty taste receptor in the absence of the test compound.
[0022] In some aspects, the human salty taste receptor comprises one alpha subunit, one beta subunit, and one gamma subunit. In other aspects, the salty taste receptor comprises one alpha subunit, one beta subunit, one gamma subunit, and one epsilon subunit. In other aspects, the salty taste receptor comprises two alpha subunits, one beta subunit, and one gamma subunit. In further aspects the salty taste receptor comprises three alpha subunits, three beta subunits, and three gamma subunits. Additional aspects include those wherein the salty taste receptor comprises one delta subunit, one beta subunit, and one gamma subunit. In other aspects, the salty taste receptor comprises two delta subunits, one beta subunit, and one gamma subunit. In still further aspects, the salty taste receptor comprises two delta subunits, two beta subunits, and two gamma subunits. In still further aspects, the salty taste receptor comprises three delta subunits, three beta subunits, and three gamma subunits.
[0023] In the method for identifying modulators of the human salty taste receptor, the delta subunit preferably comprises the amino acid sequence of SEQ ID NO:12. In some aspects, the delta receptor comprises the amino acid sequence of SEQ ID NO:9. In the method for identifying modulators of the human salty taste receptor, the method may further comprise contacting the epithelial sodium ion channel with an epithelial sodium ion channel antagonist, such as, but not limited to, chlorhexidine, amiloride, phenamil, benzamil or a homolog, analog, or derivative thereof.
[0024] In the method for identifying modulators of the human salty taste receptor, the lipid membrane may comprise at least one of phosphatidylcholine, phoshpatidylethanolamine, phostphatidylserine, phosphatidylglyine, phosphatidylinositol, sphingomyelin, cholesterol, cardiolipin, or a homolog, analog, or derivative thereof. The lipids may be organized as a liposome or lipid bilayer.
[0025] In some aspects of the method for identifying modulators of the human salty taste receptor, at least two subunits of an epithelial sodium ion channel are present in the lipid membrane at differing ratios relative to each other. The channels in the membrane preferably comprise at least one biological activity of a functional human salty taste receptor.
[0026] In the step for determining a modulation of the biological activity of the salty taste receptor, any suitable means known in the art may be used, such as, but not limited to voltage clamping, and/or measurement of an indicator dye. The method may be adapted for high throughput screening.
[0027] Compounds that modulate human salty taste perception are identified by the method of the invention and may include, for example, salty taste mimics, enhancers, modifiers, and inhibitors. The invention thus provides modulators of human salty taste perception which may further be used in compositions by admixing the compounds with a pharmaceutically acceptable carrier, or foods and beverages to modulate the salty taste perception of the food or beverage.
[0028] The invention also provides an artificial lipid membrane comprising at least one type of phospholipid and an epithelial sodium ion channel or specific ratios of epithelial sodium ion channel subunits wherein the subunits are selected from the group consisting of alpha subunits, beta subunits, gamma subunits, delta subunits, and epsilon subunits.
[0029] The artificial lipid membrane may comprise at least one phospholipid including phosphatidylcholine, phoshpatidylethanolamine, phostphatidylserine, phosphatidylglyine, phosphatidylinositol, sphingomyelin, cholesterol, cardiolipin, or a homolog, analog, or derivative thereof. The lipid membrane may be organized, for example, as a liposome or lipid bilayer.
[0030] In some aspects, the artificial lipid membrane comprises at least one epithelial ion channel comprising one alpha subunit, one beta subunit, and one gamma subunit. In other aspects, the epithelial ion channel comprises one alpha subunit, one beta subunit, one gamma subunit, and one epsilon subunit. In other aspects, the epithelial ion channel comprises two alpha subunits, one beta subunit, and one gamma subunit. In further aspects the epithelial ion channel comprises three alpha subunits, three beta subunits, and three gamma subunits. Additional aspects include those wherein the epithelial ion channel comprises one delta subunit, one beta subunit, and one gamma subunit. In other aspects, the epithelial ion channel comprises two delta subunits, one beta subunit, and one gamma subunit. In still further aspects, the epithelial ion channel comprises two delta subunits, two beta subunits, and two gamma subunits. In still further aspects, the epithelial ion channel comprises three delta subunits, three beta subunits, and three gamma subunits.
[0031] The method also provides a method for preparing such artificial lipid membrane comprising admixing a liposome comprising at least one phospholipid with an epithelial sodium ion channel or specific ratios of epithelial sodium ion channel subunits wherein the epithelial sodium ion channel or epithelial sodium ion channel subunits are dissolved in a suitable aqueous buffer comprising at least one detergent, incubating the liposome with the epithelial sodium ion channel or epithelial sodium ion channel subunit for a sufficient amount of time, and removing the at least one detergent.
[0032] The method of preparing the artificial lipid membrane may further comprise reconstituting the proteo-liposome into a planar lipid bilayer.
[0033] The invention further provides a method for identifying modulators of salty taste perception comprising: assembling at least one epithelial sodium ion channel in a lipid membrane, wherein the epithelial sodium ion channel comprises at least one of an alpha subunit, a beta subunit, a gamma subunit, a delta subunit, or an epsilon subunit; contacting the ion channel with a test compound in the presence of sodium or lithium; determining a modulation of the biological activity of the epithelial sodium ion channel in the presence of the test compound relative to the biological activity of the epithelial sodium ion channel in the absence of the test compound; and administering the test compound to a subject and determining a modulation of salty taste perception in the subject relative to the level of salty taste perception in the subject in the absence of the test compound. Preferably, the epithelial sodium ion channel comprises at least one beta subunit, at least one gamma subunit, and at least one delta subunit.
[0034] In some aspects, the epithelial ion channel comprises one alpha subunit, one beta subunit, and one gamma subunit. In other aspects, the epithelial ion channel comprises one alpha subunit, one beta subunit, one gamma subunit, and one epsilon subunit. In other aspects, the epithelial ion channel comprises two alpha subunits, one beta subunit, and one gamma subunit. In further aspects the epithelial ion channel comprises three alpha subunits, three beta subunits, and three gamma subunits. Additional aspects include those wherein the epithelial ion channel comprises one delta subunit, one beta subunit, and one gamma subunit. In other aspects, the epithelial ion channel comprises two delta subunits, one beta subunit, and one gamma subunit. In still further aspects, the epithelial ion channel comprises two delta subunits, two beta subunits, and two gamma subunits. In still further aspects, the epithelial ion channel comprises three delta subunits, three beta subunits, and three gamma subunits.
[0035] In some aspects, the delta subunit comprises the amino acid sequence of SEQ ID NO:12. In some aspects, the delta subunit comprises the amino acid sequence of SEQ ID NO:9.
[0036] In some aspects, the subject is a human.
[0037] The method permits identification of a compound that reacts in vitro with the human salty taste receptor and which is perceived by subjects as salty. The invention thus provides such compounds which may be used in compositions by admixing the compounds with a pharmaceutically acceptable carrier, or foods or beverages to modulate the salty taste perception of the food or beverage. Preferably, the compounds allow perception of salty taste, but which have a reduced effect on blood pressure as compared to salt and which have no untoward effect on the subject.
[0038] In some aspects, the compounds can be additionally screened by cell based assays for epithelial sodium channel activity.
[0039] The invention also provides kits for identifying modulators of the human salty taste receptor comprising at least one form of phospholipid; substantially purified epithelial sodium ion channel subunits comprising alpha subunits, delta subunits, beta subunits, gamma subunits, or epsilon subunits; and optionally comprising an epithelial sodium ion channel modulator, sodium or lithium, and instructions for using the kit in a method for identifying modulators of the human salty taste receptor.
[0040] The instructions may provide, for example, directions to admix the subunits in specific ratios to achieve various forms of the epithelial sodium ion channel of interest. In some aspects, at least two subunits are added to be present at differing ratios relative to each other.
[0041] The kit may contain a modulator such as, but not limited to amiloride, phenamil, benzamil, chlorhexidine, or a source of guanidinium ion.
[0042] The invention also provides a method of modulating salty taste perception (either by stimulating salty taste perception or inhibiting salty taste perception) comprising contacting a human salty taste receptor with a compound that stimulates salty taste perception wherein the salty taste receptor comprises at least one beta polypeptide subunit, at least one gamma polypeptide subunit, and at least one delta polypeptide subunit wherein said delta polypeptide subunit comprises the amino acid sequence of SEQ ID NO:12, and wherein said compound specifically interacts with said delta subunit.
[0043] In some aspects, the human salty taste receptor comprises one alpha subunit, one beta subunit, and one gamma subunit. In other aspects, the salty taste receptor comprises one alpha subunit, one beta subunit, one gamma subunit, and one epsilon subunit. In other aspects, the salty taste receptor comprises two alpha subunits, one beta subunit, and one gamma subunit. In further aspects the salty taste receptor comprises three alpha subunits, three beta subunits, and three gamma subunits. Additional aspects include those wherein the salty taste receptor comprises one delta subunit, one beta subunit, and one gamma subunit. In other aspects, the salty taste receptor comprises two delta subunits, one beta subunit, and one gamma subunit. In still further aspects, the salty taste receptor comprises two delta subunits, two beta subunits, and two gamma subunits. In still further aspects, the salty taste receptor comprises three delta subunits, three beta subunits, and three gamma subunits.
[0044] In some aspects the compound specifically interacts a portion of the delta subunit containing the amino acid sequence of SEQ ID NO:12. In some aspects, the compound binds to the portion of the delta subunit containing the amino acid sequence of SEQ ID NO:12.
BRIEF DESCRIPTION OF THE DRAWINGS
[0045] FIG. 1 shows a human taste bud stained by an ATPase histochemical procedure.
[0046] FIG. 2 shows antibody detection of the second messenger enzyme, phospholipase Cbeta2 (PLCbeta2) using an immunohistochemical procedure on human taste cells. Panel A shows the subset of cells labeled by the antibody. Panel B is a contrast image of the taste bud and of the surrounding fungiform papillae.
[0047] FIGS. 3 (a, b, c, d) shows an alignment of ENaC delta subunit sequenced from cDNA of ten individuals (labeled DENACA, DENACD, DENACE, DENACG, DENACH, DENACI, DENACJ, DENACT, DENACTV and DENACW, respectively), as compared with the GeneBank sequence of the top row (DENACGB). DENACA, DENACD, DENACE, DENACG, DENACH, DENACI, DENACJ, DENACT, DENACTV and DENACW correspond to SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26, respectively.
[0048] FIG. 4 shows amiloride inhibition of ENaC of different composition. ENaC composed of human delta, beta, gamma was less sensitive to amiloride than that composed of human alpha, beta, gamma.
[0049] FIG. 5 shows immuno-labeling of a subset of cells in a human taste bud.
[0050] FIG. 6 shows the capture of an isolated human taste bud cell by a micropipette from an aqueous suspension. The cell thus captured is placed in an RNA-preserving medium for further study.
[0051] FIG. 7 shows an early quantitative RT-PCR of a single cell tracing the amplification of partial transcripts of the ENaC subunits, alpha, beta, gamma, and delta. The result suggests a cell containing equal copies of delta, beta, and gamma, with the alpha transcript showing as a genomic control.
[0052] FIG. 8 (a, b, c, d) shows an alignment of ENaC gamma subunit sequenced from cDNA of ten individuals (labled GENACA, GENACB, GENACD, GENACE, GENACG, GENACH, GENACJ, GENACT, GENACV, and GENACW, respectively) compared with the GeneBank sequence of the top row (GENACGB). GENACA, GENACB, GENACD, GENACE, GENACG, GENACH, GENACJ, GENACT, GENACV, and GENACW correspond to SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:36, respectively.
[0053] FIG. 9 shows single channel recording of the activity of the catfish putative taste receptor for L-arginine in planar lipid bilayers. Proteoliposomes containing purified receptor protein from catfish taste epithelium are fused to planar lipid bilayers. Control records (trace shown in part A) were obtained after addition of proteolipisomes to the membrane bathing solution before addition of L-Arg. The addition of 10 μM L-Arg to the cis-side of the bilayer evoked regular periodic channel activity (trace shown in panel B, including the inset that shows the current record at an expanded scale). After several minutes of single channel recording, 100 μM D-Arg was added to the cis-side (trace shown in panel C) and activity ceased. Transmembrane potential was -100 mV. Traces shown in all panels are continuous records of that specific condition.
DETAILED DESCRIPTION
[0054] It is to be understood that this invention is not limited to particular methods, reagents, compounds, compositions, or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting.
[0055] Various terms relating to the methods and other aspects of the present invention are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definition provided herein.
[0056] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a cell" includes a combination of two or more cells, and the like.
[0057] The term "about" as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and still more preferably ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.
[0058] As used herein the "Epithelial Sodium Channel" or, as abbreviated, "ENaC," refers to a multisubunit protein that is responsible for flow of or transport of sodium ions across specific epithelium or cell membranes. ENaCs are generally composed of multiple subunits, generally α, β, γ subunits. There are also δ and ε subunits which may be in some ENaCs in specific tissues. The "salty taste receptor" as discovered herein, is a species of ENaC that is localized in taste cells and in one aspect is composed of β, γ, and δ subunits.
[0059] As used herein, "test compound" refers to any purified molecule, substantially purified molecule, molecules that are one or more components of a mixture of compounds, or a mixture of a compound with any other material that can be analyzed using the methods of the present invention. Test compounds can be organic or inorganic chemicals, or biomolecules, and all fragments, analogs, homologs, conjugates, and derivatives thereof. Biomolecules include proteins, polypeptides, nucleic acids, lipids, monosaccharides, polysaccharides, and all fragments, analogs, homologs, conjugates, and derivatives thereof. Test compounds can be of natural or synthetic origin, and can be isolated or purified from their naturally occurring sources, or can be synthesized de novo. Test compounds can be defined in terms of structure or composition, or can be undefined. The compound can be an isolated product of unknown structure, a mixture of several known products, or an undefined composition comprising one or more compounds. Examples of undefined compositions include cell and tissue extracts, growth medium in which prokaryotic, eukaryotic, and archaebacterial cells have been cultured, fermentation broths, protein expression libraries, and the like.
[0060] As used herein, the terms "modulate" means any change, increase, or decrease in the amount, quality, or effect of a particular activity or protein. "Modulators" refer to any inhibitory or activating molecules identified using in vitro and in vivo assays for, e.g., agonists, antagonists, and their homologs, including fragments, variants, and mimetics, as defined herein, that exert substantially the same biological activity as the molecule. "Inhibitors" or "antagonists" are modulating compounds that reduce, decrease, block, prevent, delay activation, inactivate, desensitize, or downregulate the biological activity or expression of a molecule or pathway of interest. "Inducers," "activators," or "agonists" are modulating compounds that increase, induce, stimulate, open, activate, facilitate, enhance activation, sensitize, or upregulate a molecule or pathway of interest. In some preferred aspects of the invention, the level of inhibition or upregulation of the expression or biological activity of a molecule or pathway of interest refers to a decrease (inhibition or downregulation) or increase (upregulation) of greater than from about 50% to about 99%, and more specifically, about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69% 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more. The inhibition or upregulation may be direct, i.e., operate on the molecule or pathway of interest itself, or indirect, i.e., operate on a molecule or pathway that affects the molecule or pathway of interest.
[0061] "Pharmaceutically acceptable carrier" refers to a medium that does not interfere with the effectiveness of the biological activity of the active ingredient(s) of a composition, and is not toxic to the subject to which it is administered.
[0062] "Ct" or "threshold cycle" refers to the PCR cycle in which a noticeable increase in reporter fluorescence above a baseline signal is initially detected.
[0063] "ΔCt" refers to the difference between the Ct of a sample assay and the Ct of a control sample. Thus, ΔCt=Ct(target)-Ct(control).
[0064] "ΔΔCt" refers to the difference between the average ΔCt value of a target sample and the average ΔCt for a corresponding calibrator sample. Thus, ΔΔCt(test sample)=AvgΔCt(test sample)-AvgΔCt(calibrator sample).
[0065] "Biological activity" as used herein refers to a measurable function of an ENaC, including but not limited to, maintenance of a sodium gradient across the membrane, changes in ion flux, changes in membrane potential, current amplitude, voltage gating, sensitivity to chlorhexidine, amiloride, or its analogs, stimulation by bretylium, novobiocin, or guanidinium ions, binding to subunit-specific monoclonal antibodies, and the like.
[0066] The present invention is based on the discovery that the human salty taste receptor is an epithelial sodium ion channel. It is thus an object of the present invention to use the precise molar ratios of the ENaC subunits and to reconstitute the ENaCs in a lipid bilayer in order to identify compounds that modulate the biological activity of the ENaCs. In particular it is an object of the present invention to use the precise molar ratios of the salty taste receptor subunits to reconstitute the salty taste receptor in the a lipid bilayer in order to identify compounds that modulate the biological activity of the salty taste receptor and to identify compounds that modulate salty taste perception in human beings. Without intending to be limited to any particular theory or mechanism of action, it is believed that a passive influx of sodium ions through epithelial sodium channels in certain taste receptor cells causes a change in intracellular ion balance leading to a depolarization, ultimately resulting in neurotransmitter release, which in turn produces a perception of salty taste.
[0067] In one aspect, the invention provides assays to identify compounds that bind and/or modulate the human salty taste receptor. The methods comprise assembling at least one epithelial sodium ion channel in a lipid membrane, wherein the epithelial sodium ion channel comprises an alpha, beta, gamma, or delta subunit, contacting the at least one ion channel with a test compound in the presence of sodium or lithium, and determining a modulation of the biological activity of the at least one epithelial sodium ion channel in the presence of the test compound relative to the biological activity of the at least one subunit in the absence of the test compound.
[0068] Where the biological activity of the sample containing the test compound is higher than the activity in the sample lacking the test compound, the compound is an agonist. If the activity of the sample containing the test compound is lower than the activity in the sample lacking the test compound, the compound is an antagonist.
[0069] Epithelial sodium ion channels are heteromultimeric complexes that are comprised of different subunits. Various subunits of ENaC have been identified, and include, without limitation, the alpha subunit, the beta subunit, the gamma subunit, the delta subunit, and the epsilon subunit. The ENaC subunits may derived from any species, however, mammalian ENaC subunits are preferred and the most preferred species is human. Examples of nucleic acid sequences encoding human ENaC subunits and the deduced amino acid sequences are provided herein. Other subunits with amino acid sequences that are substantially homologous or which represent isoforms of the subunit proteins may be used in practicing the invention Amino acid sequences that are "substantially homologous" are at least protein sequences that are from about 80% to about 100% identical to the sequence provided herein for the subunit sequence. More preferably, the sequences are about 85% to about 100% identical. Most preferably, the sequences are about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the reference sequence provided herein for the subunit.
[0070] Representative nucleotide sequences encoding human alpha subunit, human beta subunit, human gamma subunit, and human delta subunit are provided as SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, and SEQ ID NO:7, respectively. The deduced amino acid sequences for human alpha subunit, human beta subunit, human gamma subunit, and human delta subunit are provided as SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8, respectively. In a preferred aspect for the salty taste receptor, the delta subunit comprises a cysteine at position 532 with respect to SEQ ID NO:8. The delta receptor with Cys532 is shown in SEQ ID NO:9. Such substitution may arise due to a alteration in the triplet codon from tac to tgc (with respect to that shown in SEQ ID NO:7) and results in a change from tyrosine (Tyr) to cysteine (Cys).
[0071] In some aspects, the ENaC is comprised of at least one alpha subunit, at least one beta subunit, and at least one gamma subunit (e.g., (α)1(β)1(γ) 1). In other aspects, the ENaC is comprised of at least one alpha subunit, at least one beta subunit, at least one gamma subunit, and at least one delta subunit. In other aspects, the ENaC is composed of two alpha subunits, one beta subunit, one gamma subunit (α2βγ). In other aspects, the ENaC is composed of three alpha subunits, three beta subunits, and three gamma subunits (α)3(β)3(γ)3). In another aspect, the ENaC comprises an epsilon subunit and at least one other subunit such as an alpha subunit, beta subunit, delta subunit, gamma subunit, or combinations thereof. In still other aspects the ENaC comprises a plurality of beta subunits. In a preferred aspect, the ENaC is comprised of at least one beta subunit, at least one gamma subunit, and at least one delta subunit (the salty taste receptor). The most preferred aspect is an ENaC comprising at least one beta, at least one gamma, and at least one delta subunit (e.g., (β)1(γ)1)(δ)1 in which the delta subunit contains Cys532.
[0072] The various subunits can be present in the ENaC in different ratios relative to other subunits. The observed variation may relate to which tissue the particular ENaC of interest is expressed in. For example, but not by way of limitation, an ENaC can be comprised of two alpha, one beta, and one gamma subunit. Thus, in certain aspects of the invention, the ENaC assembled into a lipid membrane is comprised of at least two subunits that are present in different ratios relative to the other subunits. In other aspects, the ENaC is comprised of at least two subunits that are present in the same ratio relative to the other subunits. The ratios of the ENaC subunits may also vary depending on the tissue in which the ENaCs of interest are expressed. Further, there may be important sequence variability in the form of each subunit expressed in various tissues. For example, but not by way of limitation, the delta subunit of ENaC expressed in the salty taste receptor preferably has a cysteine in the putative amiloride binding site of delta at position 532 of SEQ ID NO:8 (which encodes human delta from kidney). Human kidney delta has a tyrosine at this position. Thus, when expressing a human salty taste receptor, it is preferred to use a delta with the putative amiloride binding site of MGSLCSLWFGA (SEQ ID NO:12) which includes CYS532. As this motif is at least a putative site for amiloride binding, other compounds that modulate the human salty taste receptor may also bind to this site.
[0073] In certain aspects of the invention, the lipid membranes produced with the ENaC subunits in them contain ENaC subunits that form the salty taste receptor. These salty taste receptors include at least one beta, at least one gamma, and at least one delta subunit. In preferred aspects, the delta subunit comprises Cys532. In other aspects, the ENaC contains subunits selected from alpha, beta, gamma, delta, and epsilon. In some aspects, the ENaC is composed of at least one alpha, at least one beta, and at least one gamma. In other aspects, the ENaC comprises at least one epsilon subunit.
[0074] The ENaC or the various subunits that are to be assembled into the lipid membrane can be obtained from any source suitable in the art. For example, an ENaC or any subunit thereof can be freshly isolated from any cell that expresses and ENaC, including cell lines and stable cell lines. For example, but not by way of limitation, ENaC are expressed in neural tissue, the pancreas, testes, ovaries, tongue, colon, kidneys, lungs, sweat glands, and the like. In some aspects, the ENaC for salty taste perception is isolated from the papillae of the tongue. In other aspects, an ENaC or any subunit thereof can be recombinantly expressed, purified and used to reconstitute a lipid membrane to form functional ENaCs.
[0075] In certain aspects, each subunit of the ENaC is separately expressed in a recombinant expression system such as, but not limited to bacterial cells, Spodoptera frugiperda cells, mammalian cells, and frog oocytes. The expressed protein is purified by standard biochemical means as is well-known in the art. Alternatively, expressed protein may be immunopurified using immobilized antibodies that specifically bind the ENaC subunits. Methods for purifying proteins by immunoaffinity (using antibodies that specifically bind the subunit or ENaC of interest). In other aspects, the ENaC subunits are expressed as a fusion protein with a polypeptide that allows for rapid purification and subsequent cleavage from the expressed protein. Such purification systems include, but are not limited to the pGEX system (glutathione-S-transferase fusion proteins) and multi-histidine fusion proteins (for nickel binding affinity purification). These and other types of purification are described in numerous references and are well-known to those of skill in the art. In certain preferred aspects, the ENaC subunits are expressed simultaneously using a baculovirus system and Spodoptera frugiperda cells and membrane fractions are prepared as described in Rao, U.S. et al. (2002) "Activation of Large Conductance Sodium Channels Upon Expression of Amiloride-Sensitive Sodium Channel in SF9 Insect Cells" J. Biol. Chem. 277(7):4900-4905.
[0076] In certain aspects, the subunits of the ENaC are substantially purified prior to incorporation into the membrane. As used herein, "substantially purified" refers to subunits that are at least 80% free of contaminating material (e.g., proteins, polysaccharides, and lipids) derived from the cells from which they are obtained. Preferably, the subunits are at least about 85% free of contaminating material. More preferably, the subunits are at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more free of contaminating material.
[0077] To recreate a particular ENaC which may be present in a tissue in the artificial membranes of the invention, the ratio of the subunits present in the ENaC may be determined by quantitative PCR. As the ratio of protein subunits of a multimeric receptor often correlates to the amount of mRNA produced in a cell for the given receptor, quantitative PCR can provide an efficient means of determining the ratio of mRNA present. Protocols for performing quantitative PCR are well known in the art. Further, given the sequences of the ENaCs provided herein and the knowledge in the art and software available for selecting PCR oligonucleotide primers that can specifically and reliably amplify messages for particular genes, one of skill in the art may easily and routinely perform quantitative PCR on tissue samples and determine the identity and ratio of the subunits that form a particular ENaC. Assays for determining the relative amounts of mRNA are well known in the art. Once the ratio of mRNA is determined, one may extrapolate the amount of protein of each subunit that must be added to the membrane to provide the appropriate stoichiometric amounts of protein to form biologically active ENaCs.
[0078] The concentration of affinity-purified protein can be determined by measuring the total nitrogen content of the protein eluate and comparing the nitrogen content with the total protein content of the eluate. Nitrogen content can be determined by any means suitable in the art, such as the well-known Kjeldahl Nitrogen Method. Protein concentration can be determined, for example, by spectrophotometry whereby a protein sample is analyzed for its absorption of light at 280 nm to derive an absorption coefficient. Any means known in the art for assessing concentration and/or purity of protein may be used.
[0079] The invention thus provides artificial membrane systems containing substantially purified ENaC protein subunits that assemble into functional ENaCs. Specifically, the invention provides artificial membrane systems containing substantially purified human salty taste receptor. These membrane systems permit analysis of ENaCs, including, but not limited to the salty taste receptor apart from contaminating proteins such as endogenous ENaCs. The invention permits the assembly of ENaCs in which the subunits are added at known ratios to permit the assembly of precise ratios of selected subunits. The lipid membrane can comprise any combination of lipids. Non-limiting examples of suitable lipids include phosphatidylcholine, phoshpatidylethanolamine, phostphatidylserine, phosphatidylglyine, phosphatidylinositol, sphingomyelin, cholesterol, cardiolipin, or a homolog, analog, or derivative thereof. Phospholipids are preferred, and can be obtained from any source suitable in the art. For example, the phospholipids can be extracted from a cell, or can be synthetic phospholipids, which are commercially available.
[0080] The lipid membrane can be in any conformation or phase, including without limitation, liposomes, a lipid bilayer, or the hexagonal phase. Liposomes and lipid bilayers are particularly preferred.
[0081] The effect of the test compound on the biological activity of the ENaC an be determined by any means suitable in the art. The test compound can be assessed at multiple concentrations. In some aspects, the test compound is assessed for its ability to modulate at least one biological activity of the ENaC. In preferred aspects, the ENaC is the salty taste receptor.
[0082] The biological activity of the ENaC can be determined by measuring the current of ENaC assembled in the lipid membrane. Voltage clamping is one preferable technique to measure ENaC current. Voltage clamp techniques are well known in the art. (Nagel, G et al. (2005) J. Physiol. 564(Pt 3):671-82; Staruschenko, A et al. (2004) J. Biol. Chem. 279:27729-34; Tong, Q et al. (2004) J. Biol. Chem. 279:22654-63; Sheng, S et al. (2000) J. Biol. Chem. 275:8572-81). The following parameters can be measured using a voltage clamp: single channel conductance, channel open time, voltage dependence, blockade induced by application of a particular compound, and activation induced by application of a particular compound. Other suitable techniques for measuring the biological activity of ENaCs include flux assays, patch clamping, voltage-sensitive dyes, and ion-sensitive dyes. Preferably, ENaC activity is measured by membrane electrophysiology or by assessing the change in fluorescence of a membrane potential dye in response to sodium or lithium, or analogs thereof (e.g., isotopes). All such assays are well known in the art. (Gill, S et al. (2003) Assay Drug Dev. Technol. 1:709-17, flux assay; Caldwell, R A et al. (2005) Am. J. Physiol. Lung Cell Mol. Physiol. 288:L813-9, patch clamp). A variety of voltage sensitive dyes are commercially available, including without limitation styryl dyes, oxonol dyes, and merocyanine-rhodanine dyes. Selection of the appropriate voltage sensitive dye is within the relevant skill in the art. Similarly, a variety of ion sensitive dyes are commercially available, including single excitation dyes, dual excitation ratiometric dyes, and dual emission ratiometric dyes.
[0083] The salty taste receptor is responsive to sodium and lithium ions. However, unlike other ENaCs, the human salty taste receptor is not sensitive to amiloride. Thus, amiloride should not inhibit or stimulate the salty taste receptor ENaC. Conversely, chlorhexidine acts as an inhibitor of the salty taste response in humans, and may be used in assays to identify salty taste modifiers. One may assess specificity of stimulation of the salty taste receptor with test compounds by showing that the effect is inhibited by chlorhexidine. Moreover, test compounds that can overcome the effect of chlorhexidine (and stimulate the salty taste receptors in the membrane systems of the invention) are strong salty taste enhancers. Basic compounds containing guanidinium ions as well as certain amines act as salty taste enhancers. These include guanidine, arginine, and homoarginine. Both L- and D-arginine are equally effective. While not wishing to be bound by any particular theory of operation, this lack of enantiomeric specificity suggests that the primary enhancing effect derives from a compact, basic moiety, in this case the guanidinium ion. Thus, salty taste receptors in the membrane system of the invention may be stimulated by contacting them with a source of guanidinium ions. It may be assumed that these enhancing compounds interact directly with the human salty taste ion channel, since most sodium channel blockers and enhancers contain guanidinium groups that interact with acidic moieties inside the channel pore lumen. Thus, the molecular mechanisms of human salty taste share selected functional features in common with known sodium channels but also have unique pharmacological attributes.
[0084] Amiloride and amiloride derivatives (e.g., phenamil, benzamil and the like) may be useful in assessing other ENaCs, such as those containing an alpha subunit. Amiloride and derivatives may also be used in assays to inhibit the background (endogenous ENaCs) if the purity of the subunit preparations is low such that host cell ENaCs are contaminating the preparation. Thus, in some aspects of the invention, the methods further comprise contacting the ENaC with a sodium ion channel antagonist. Such antagonists are well-known to those of skill in the art. Preferably, the antagonist is amiloride, chlorhexidine, or homologs, analogs, or derivatives thereof.
[0085] The invention also includes within its scope high-throughput screening assays to identify compounds that modulate the biological activity of the salty taste receptor. High-throughput screening assays permit screening of large numbers of test compounds in an efficient manner. For example, but not by way of limitation, lipid membranes comprising an assembled ENaC can be dispersed throughout a multi-well plate such as a 96-well microtiter plate. Each well of the microtiter plate can be used to run a separate assay against a candidate modulator. A microtiter plate permits screening of multiple concentrations of a test compound, multiple test compounds, alone or in combination with other test compounds, and multiple ENaCs, including ENaCs with varying ratios of subdomains as described and exemplified herein under identical assay conditions. In other aspects of the invention, planar lipid bilayers containing the ENaC of interest is contacted with a test compound and a measurement is taken. The solution on one or both sides of the planar bilayer is changed and the bilayer is contacted with a second test compound. This may be continuously used as a high-throughput assay. The assays may take place in the presence of additional agonists or antagonists. Data obtained for the test compounds are compared with measurements taken in the presence of known agonists or antagonists and/or to control samples (such as a non-stimulatory/non-inhibitory medium).
[0086] Serial assays may be performed to narrow down the pool of test compounds that act as salty taste modifiers. For example, the in vitro assays of the invention may be combined with cell-based assays as a secondary or confirming screening step. Such assays have been described, for example, in published U.S. Patent Application 2005/0059094 to Servant et al.
[0087] An additional aspect of the invention features methods for identifying compounds that modulate salty taste perception in a subject by a combination of in vitro and in vivo screening assays. In one aspect, a test compound is first screened in vitro to determine its modulatory effect on an epithelial sodium ion channel, and then screened further in vivo to determine if the compound can modulate, preferably enhance, salty taste perception in a subject.
[0088] In one aspect, the in vitro screening assay comprises identifying modulators of the human salty taste receptor comprising contacting a test compound with at least one ENaC and determining a decrease in the biological activity of the ENaC in the presence of the test compound relative to the biological activity of the ENaC in the absence of the test compound. This aspect can be practiced according to the details described herein. In one aspect, the in vivo screening assay comprises identifying compounds that enhance salty taste perception in a subject comprising administering a test compound to the subject and determining whether salty taste perception is enhanced in the subject relative to the level of salty taste perception by the subject in the absence of the test compound.
[0089] For in vivo screening, subjects can be recruited via an Institutional Review Board-approved method such as general advertisement in print media. Prior to entering the study, each subject provides informed consent. The participants can be requested to refrain from eating, drinking or chewing gum for at least one hour prior to testing. Subjects can be paid to participate in the study.
[0090] Experimental solutions containing a candidate test compound to be administered to study subjects can be presented in the form of a binary mixture such as the compound and an inorganic salt such as NaCl. Preliminary experiments can be carried out to establish an appropriate concentration range for the test compound and inorganic salts. For example, four concentrations of the test compound are used with four concentrations of the inorganic salts. Aqueous solutions can be prepared to encompass all possible combinations of the concentrations of the test compound with the inorganic salts.
[0091] To assess the salty taste amplifying properties of a given stimulus, any means suitable in the art can be used. One non-limiting example of such means is the method of magnitude estimation. Magnitude estimation measures ratings of the perceived intensities of saltiness from a sample. Subjects participating in saltiness assessments can be instructed to rate the saltiness or relative saltiness of a solution. Each solution can be sampled by the subject once, twice, three times, or more.
[0092] Prior to sampling a test solution, subjects can be instructed to rinse their mouth. For example, subjects can be instructed to rinse with and expectorate water four times, preferably within a short duration of time such as period of approximately two minutes. Test samples and inorganic salt solutions can then be administered to the subjects, preferably in random order, and without replacement. For example, solutions can be prepared in polystyrene medicine cups (Dynarex, N.Y.) in 10 ml aliquots, and administered to the subjects. The subject can be instructed to rate the relative saltiness of the solution, and the relative saltiness ratings for each solution can be arithmetically averaged to yield single ratings of saltiness.
[0093] Magnitude estimation may not reveal differences due to variations in subject number use. To eliminate the variance produced by idiosyncratic number usage in the magnitude estimation task, the saltiness ratings can be standardized to the grand arithmetic mean of the saltiness ratings of NaCl alone in water (averaged across all NaCl concentrations). Each subject's mean saltiness rating can be divided into the grand saltiness mean, and the quotient can be used as the multiplicative standardization factor for that individual's saltiness rating. This procedure equates mean saltiness ratings of NaCl in water across subjects.
[0094] Analysis of variance (ANOVA) can be conducted on the standardized repeated measures data from the magnitude estimation, and post-hoc pairwise comparisons can be conducted with Tukey's honest significant difference (HSD) analysis.
[0095] An alternative to magnitude estimation is a forced-ranking procedure, wherein a series of two-alternative forced-choice pairings are used to rank the saltiness of aqueous solutions of NaCl in the presence or absence of a test compound putative enhancer. In this procedure, subjects can be instructed to taste half of the first solution (for example, 5 ml or 10 ml solution) of the first pair of samples for three seconds and expectorate. Subjects then rinse twice and taste half of the second sample, expectorate, rinse twice and taste the remainder of the two solutions using the same procedure. After tasting both solutions twice, subjects can be asked to indicate which solution they thought was saltier. If they report that neither solution seemed saltier, subjects can be asked to guess (forced-choice). The procedure can repeated for all samples.
[0096] The saltiness rankings can be calculated based on the number of times a particular solution is chosen as being saltier than all other solutions using the Friedman analysis of pairwise rankings. The Tukey HSD can be calculated to determine if the differences between individual rankings are significant.
[0097] Compounds identified by any of the foregoing inventive screening methods are contemplated to be within the scope of this invention. Such compounds are preferably agonists of ENaCs, more preferably agonists of the human salty taste receptor, and even more preferably are enhancers of human salty taste receptors. Such compounds may be formulated as a nutraceutical or pharmaceutical composition by admixing such compound in an amount effective to enhance salty taste perception in the subject to which it is administered and a pharmaceutically or nutraceutically acceptable carrier, as described herein.
[0098] It is an object of the invention to use the assays to identify compounds that are perceived as salty, as well as to identify compounds that enhance salty taste (such that a reduced amount of sodium or lithium is perceived as a higher concentration of sodium or lithium). The invention enables the screening of libraries of compounds including natural or synthetic molecules including, but not limited to proteins, peptides, oligonucleotides, polynucleotides, polysaccharides, lipids, small organic molecules, and the like, for their ability to act as salt substitutes, salty taste enhancers, or salty taste inhibitors. The invention includes salt substitutes, salty taste enhancers, and salty taste inhibitors identified by the methods of the invention.
[0099] Also featured in accordance with the present invention are artificial lipid membranes and methods for preparing the same. The artificial lipid membranes comprise at least one lipid and an assembled ENaC or at least one subunit of an ENaC. In preferred aspects, the ENaC is a salty taste receptor. The lipid membrane can be comprised of any suitable lipid, and are preferably comprised of phospholipids. Suitable lipids include, without limitation, phosphatidylcholine, phoshpatidylethanolamine, phostphatidylserine, phosphatidylglyine, phosphatidylinositol, sphingomyelin, cholesterol, cardiolipin, or a homolog, analog, or derivative thereof, and these can be obtained from any source suitable in the art. The lipid membrane can be in any conformation, and preferably is a liposome or lipid bilayer.
[0100] In one aspect, an artificial lipid membrane is prepared by admixing a liposome that comprises at least one phospholipid with an ENaC or a particular subunit or subunits of an ENaC that has dissolved in a suitable aqueous buffer. The aqueous buffer comprises at least one detergent. Suitable detergents are well known in the art, and include without limitation, Tween, Triton, CHAPS, sodium cholate, and octyl-glucoside. After mixing the phospholipids and ENaC or subunits thereof, the mixture is allowed to incubate for several minutes, preferably at least about 20 minutes, to permit assembly of the ENaC into a lipid membrane. Following the incubation, the detergent is removed according to any means suitable in the art, such as those described and exemplified herein. Other methods known in the art of preparing lipids and liposomes containing proteins may be used to produce the lipids and liposomes containing the ENaC subunits.
[0101] In some aspects, the ENaC is assembled into a liposome. The liposome can be converted into a planar lipid bilayer by use of techniques that are well known and routine in the art, including those that are described and exemplified herein. In some aspects, the liposomes contain a substance other than found in the surrounding milieu. For example, but not by way of limitation, the liposomes may contain a fluorescent voltage-sensitive or membrane potential dye that is responsive to sodium or lithium, to indicate a change in sodium content as a marker of sodium flow upon stimulation with a test compound.
[0102] The invention also features kits for identifying modulators of the human salty taste receptor. The kits comprise at least one phospholipid, an isolated epithelial sodium ion channel subunit(s), and optionally a source of sodium and/or lithium ions, and instructions for using the kit in a method for identifying modulators of the human salty taste receptor. In some aspects, the kits optionally comprise an epithelial sodium ion channel antagonist and/or agonist.
[0103] The invention provides a method for modulating salty taste perception in a subject by contacting a salty taste receptor with a compound that specifically interacts with the putative amiloride-sensitive region of the delta subunit that contains Cys532. In human subjects, this delta subunit comprises the amino acid sequence of SEQ ID NO:12. The modulators may enhance or inhibit salty taste perception by either stimulating the receptor or blocking the receptor. The compounds may interact with the delta receptor by binding to the receptor, preferably in the putative amiloride sensitive region having the amino acid sequence of SEQ ID NO:12.
[0104] Using computer programs for rational-based drug design that are available in the art, molecular modeling may be performed based on the primary amino acid sequence data available herein and knowledge in the art as to tertiary structure of ion channels to provide a three dimensional model of the human salty taste receptor. Such modeling permits the rational selection of candidate compounds that will interact with specific modulatory sites including, as example, the putative amiloride binding site of the delta subunit, motif SEQ ID NO:12. These compounds, or classes of compounds, will act as salty taste modifiers. Compounds that interact with these regions (e.g., delta subunit SEQ ID NO:12) are useful as modifiers of salty taste perception. Thus, the data presented herein provide a structural-functional relationship between the subunits comprising the salty taste receptor and the areas of the subunits that are likely involved in salty taste perception.
[0105] The following actual and prophetic examples are provided to describe the invention in more detail. They are intended to illustrate, not to limit the invention.
Example 1
Procedure for Obtaining Human Fungiform Papillae and Taste Cells
[0106] Human fungiform papillae containing taste buds are routinely obtained from the anterior dorsal surface of the tongues of volunteers by a minor surgical biopsy procedure carried out under local anesthesia. The general procedure is described in Spielman, A I et al. Collection of taste tissue from mammals. Experimental Cell Biology of Taste and Olfaction. Spielman A I and Brand J G eds. CRC Press, Boca Raton, Fla., pp 25-32. Volunteers give informed consent. This procedure has been reviewed and approved by an Institutional Review Board. The excised papillae can be subsequently used either for RNA extraction, immunohistochemistry or in situ hybridization, or in a procedure that results in a suspension of isolated taste cells.
[0107] RNA extraction, histochemistry an in situ analysis. When used for total RNA extraction, papillae are immediately subjected to a standard extraction procedure using TRIzol® reagent (Invitrogen, Carlsbad, Calif.). The RNA extract is treated with DNase to remove most genomic DNA. Any DNA remaining could otherwise yield false positive results in subsequent steps where the use of intron-spanning primers is not possible. Genomic material, however, is useful in quantitative reverse trancriptase polymerase chain reaction (QRT-PCR) because the single copy of the genomic DNA signals the point of highest sensitivity of the PCR, and provides thereby a convenient end-point for the procedure. Reverse transcription is then performed on the RNA to yield a DNA copy of the RNA, known as complementary DNA or cDNA. This cDNA will used as the substrate in the polymerase chain reaction.
[0108] Because the fungiform papillae RNA and subsequent cDNA are generally of high quality, the entire coding sequence or open reading frame (ORF) of the protein under study can be immediately amplified. The oligo-nucleotide primers used to effect this amplification are designed based on the published sequence of the same or similar protein annotated in GenBank. The PCR reaction products can be analyzed by agarose gel electrophoresis. This procedure is often used to obtain the entire coding sequence of a gene known to be expressed in taste bud cells, the full sequence of which cannot be obtained readily from single cell analysis.
[0109] The excised papillae may also be used for general or immunohistochemical, or in situ hybridization analysis. Various techniques and procedures are available and can be used to fix and protect the tissue. As an example, FIG. 1 shows a human taste bud stained by an ATPase histochemical procedure. FIG. 2 shows antibody detection of the second messenger enzyme, phospholipase Cbeta2 (PLCbeta2) using an immunohistochemical procedure on human taste cells. The procedure is as follows: Histological sections (8 to 10 microns) of fungiform papillae were washed three times in 1×PBS for 10 minutes, placed in blocking buffer at room temperature for 4-18 hours. Blocking buffer was removed and primary antibody (rabbit anti-PLCbeta2) was added in three concentrations (1:50, 1:100, and 1:200 in buffer). The primary antibody solution was removed and the slides were washed three times in PBS. The first wash drained immediately while the subsequent washes were incubated for 10-20 minutes each. Excess fluid was removed and a the secondary antibody solution (CY3-labeled goat anti-rabbit, 1:1000) was added to the sections and the slides were incubated at room temperature for 45-120 minutes. The slides were washed three times in PBS. The first wash drained immediately while the subsequent washes were incubated for 10-20 minutes each. The excess fluid was drained, but slides were allowed to remain wet. Coverslips were placed on the slides and the slides were examined under a fluorescence microscope.
[0110] RT PCR for Identifying ENaC Subunits and Sequencing the Same.
[0111] Extraction of total RNA from biopsied fungiform papillae is carried out as described above, without DNase treatment, followed by synthesis of first-strand cDNA. Amplification of ENaC subunits (no more than 500 bp in size) can be performed with the PCR Core System I reagent kit (Promega Corp., Madison Wis.) using primers as above.
[0112] If a product of apparently the correct size is obtained, this product is excised from the gel and purified. The product is then ligated into a plasmid vector to yield a recombinant plasmid which has the gene for the coding sequence of the protein (e.g., ENaC 6) inserted into it. The recombinant plasmid is used to transform bacterial cells which, when provided with an appropriate growth medium, produce large amounts of plasmid. Purification of the bacterial culture yields the recombinant plasmid in a pure form, which enables one to get the sequence of the protein gene from human fungiform papillae. Finally, a bioinformatic analysis of the sequence, using the BLAST program confirms that the correct sequence has indeed been obtained.
[0113] Using this procedure, evidence was found for transcripts of four ENaC subunit types in human fungiform papillae. These subunits are the alpha, beta, gamma, and delta ENaC subunits. The complete ORF of the alpha subunit was rarely observed, but the complete ORF of the other subunits was nearly always observed. Surprisingly, it was discovered that the delta subunit of ENaC is present in human fungiform papillae.
Example 2
[0114] Identification of the Human Salty Taste Receptor and the Importance of the Delta Subunit
[0115] In accordance with the present invention, the delta subunit of the ENaC in the fungiform (taste) papillae of humans. The clones in which the subunit was detected were from pooled cDNA from 3 individuals who agreed to undergo a biopsy procedure to remove several fungiform papillae from the anterior dorsal surface of the tongue.
[0116] Characteristics of the Delta Subunit.
[0117] The delta subunit of the epithelial sodium channel was detected in the fungiform papillae from thirteen individuals by RT-PCR. The detected fragments were amplified by PCR and subcloned. The polynucleotide encoding delta subunit from these thirteen individuals was then fully sequenced. It was determined that the human delta subunit from fungiform papillae differed from human delta subunit cloned from kidney in the putative amiloride binding site. The putative amiloride binding site contains a tyrosine at amino acid 532 in delta subunit from kidney (SEQ ID NO:8), but amino acid 532 in delta from fungiform papillae was cysteines in each of the thirteen samples sequenced (SEQ ID NO:9):
TABLE-US-00001 TABLE 1 Sequence of putative amiloride Source of delta binding site Kidney delta: MGSLYSLWFGA (SEQ ID NO: 11) Taste delta# 1: MGSLCSLWFGA (SEQ ID NO: 12) Taste delta# 2: MGSLCSLWFGA (SEQ ID NO: 12) Taste delta# 3: MGSLCSLWFGA (SEQ ID NO: 12)
[0118] FIG. 3 shows an amino acid sequence alignment of 11 delta subunits, where the first sequence is the GenBank sequence with the other 10 sequences being from 10 different individuals. At position 180, a possible polymorphism (R to P) is indicated. Other positions indicating possible polymorphisms are with positions 278 (F to I), 355 (S to R), 389 (E to Q), and 566 (R to H). Position 566 has also been implicated in amiloride binding. Without intending to be limited to any particular theory or mechanism of action, the polymorphisms may play a role in sensitivity to salty stimuli or may play a role in sensitivity to taste modulators.
[0119] The Y to C change at position 532 is significant as it may help explain why rat salty taste receptors are apparently amiloride sensitive while human salty taste receptors are not. As the rat's delta ENaC subunit is a psuedogene, it is not expressed. It is believed that the amiloride-sensitive alpha subunit functions as part of the salty receptor in rat. Although this substitution does not significantly alter the receptor sensitivity and specificity, the pharmacology of the channel is altered.
[0120] While the delta subunit is amiloride sensitive, it is less so than the alpha (FIG. 4). Thus, if the human salty taste receptor ENaC contained the usual form of delta, it too should show amiloride sensitivity. However, the putative amiloride binding site in delta from human taste cells contains a non-conservative substitution and may therefore have a different sensitivity to amiloride than delta subunit in kidney. Without intending to be limited to any particular theory or mechanism of action, it delta shows less sensitivity, this observation potentially can be interpreted to mean that delta is in the human salty taste receptor, particularly because amiloride cannot cross tight junctions. Because of the differences between rat and human, the rat is probably not a good model for salty taste perception in humans.
[0121] Cellular Specificity of the Human Fungiform Delta ENaC Subunit.
[0122] A human taste bud is shown in FIG. 1, wherein an 8 micron section of a human fungiform papilla is stained by an ATPase histochemical procedure. The question now became whether some, all, or none of these taste cells expressed delta ENaC. To view only those cells expressing the delta subunit, an antibody to the delta form of human ENaC was raised in rabbits. A representative photograph is shown in FIG. 5. The slide shows tissue specific labeling on a subset of cells within a human taste bud. The implication is that the human salty taste receptor is an ENaC composed of a multimer of delta, beta, and gamma subunits or of subunits alpha, delta, beta, and gamma This specificity of delta in the taste cells accompanied by a notable dearth of full-length alpha in these same buds, suggested that the human salty taste receptor is a delta-containing ENaC and not simply an alpha, beta, gamma ENaC as suggested by others.
[0123] Isolation of Human Taste Bud Cells.
[0124] A suspension of single isolated taste bud cells was prepared from human fungiform papillae by incubation of biopsied papillae in a collagenase-based enzymatic procedure, followed by washing of the papillae to effectively removed enzyme, then trituration of same through a glass pipette. The resulting suspension was enriched for cells of the taste bud. Individual cells were captured using a glass micropipette (See FIG. 6) and individually placed into a tube containing 2 to 10 μl of RNAlater.
[0125] The Delta Subunit is Located in Taste Bud Cells.
[0126] cDNA was derived from 7 human fungiform taste cells that were individually isolated and captured, as described above, and then pooled. A product of the correct size (˜500 bp) was noted and its identity as a human ENaC delta subunit was confirmed by sequencing. Using this PCR procedure of identifying overlapping segments of the ORF of delta ENaC, the complete ORF of taste cell delta ENaC was obtained.
[0127] Single cell RT-PCR using nested primers was also performed, and revealed that two out of twelve human taste bud cells tested provided strong evidence for expression of delta, beta, and gamma subunits, without expressing full-length alpha (data not shown). One early single cell Q-RT PCR revealed no message for the alpha subunit but approximately equal numbers of message copies for delta, gamma and beta (FIG. 7).
[0128] Using calcium imaging on a preparation of isolated taste cells, it is possible to identify those individual cells that are activated by sodium chloride. These cells are captured and their contents analyzed by Q-SC-RT-PCR. In a group of 30 salt sensitive cells, the primary expressed subunit was determined to be delta. Eight expressed delta, beta, and gamma, while 7 expressed delta, alpha, beta, and gamma.
[0129] An alignment of the amino acid sequences of the 10 gamma subunits sequenced from taste cells as compared to the GenBank sequence for gamma is shown in FIG. 8.
[0130] Having identified the salty receptor as delta, beta, gamma or delta, alpha, beta, gamma, each subunit will be expressed and reconstituted into lipid bilayers for analyses, as provided by the examples below.
Example 3
Preparation of Liposomes and Artificial Lipid Bilayers
[0131] This example demonstrates the techniques that are readily practiced to solubilize an abundant receptor from its membrane milieu, purify the receptor, and reconstitute the receptor in an artificial lipid membrane such as a lipid bilayer. Such membranes serve as an artificial biological membrane in which the receptor resumes its native conformation and can be studied in detail and in isolation, e.g., without interference from other proteins or the metabolic whimsy of a living cell.
[0132] This example, in part, describes the extraction, purification, and membrane-reconstitution of a taste receptor for L-arginine (L-arg) from catfish. The methods, which are published and described in Grosvenor, W et al. (2004) BMC Neuroscience 5:25, can be readily adapted for reconstitution of ENaC, ENaC subunits, and salty taste receptors in lipid membranes as described below. The catfish has served as a model for taste receptor studies because the receptors are very sensitive to certain amino acids. One such amino acid is L-arg. Like the ENaC, the taste receptor for L-arg in catfish is an ion channel. Parallel approaches are utilized in solubilizing the L-arg and ENaC-type receptors. The receptors differ primarily by origin--the L-arg receptor is purified from catfish taste tissue and the ENaC subunits are synthesized by a heterologous cell culture expression system.
[0133] Liposome Generation.
[0134] Liposomes are used to carry the solubilized receptor to the bilayer construct. The major challenge to studying a membrane-soluble protein is developing a procedure to move the protein from its native membrane or synthesis end point to an artificial lipid bilayer. Solubilization usually uses detergent and this detergent must be removed to avoid damage to the bilayer. The liposome performs this transfer by taking up the protein from the detergent system and giving it up to the bilayer.
[0135] Liposomes are prepared by adding 5 mg of the mixture of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine: 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPE:DOPC) in a 2:1 ratio in 0.5 ml of chloroform to a round bottom flask. The flask is rotated for 30-40 min at 4° C. After evaporation of the chloroform, a thin layer of lipid is formed to which 2 ml of buffer solution (300 mM NaCl, 50 mM Tris, pH=7) is added. After addition of the buffer, the flask is bath-sonicated 3 times with 3-5 min pulses to induce liposome formation. Alternatively, the probe sonicator can be pulsed for only 30-40 sec.
[0136] Dissolution of the L-ArgR into Liposomes and Pharmacology of L-ArgR in a Lipid Membrane.
[0137] An amount (0.01 to 0.5 mg) of the L-ArgR dissolved in 100 mM NaCl/50 mM Tris, pH=7.1 containing one of several detergents such as Tween, CHAPS, Triton, Sodium Cholate, or/and Octyl-glucoside is added to the liposomes. The ratio of protein to lipid (mass:mass) for measuring channel activity as single cannel events is 1:2000-5000. The ratio for measuring macroscopic properties is in the range of 1:50-100. The protein-lipid mixture is incubated for 20-30 mins.
[0138] The properties of the L-ArgR as measured in the bilayer are very similar to those observed through taste nerve recordings from the animal, when the L-ArgR is in its native state. For example, the taste nerve recordings indicate that the sensitivity of the native receptor is in the tenths of uM of L-arg, while D-arg inhibits the receptor. FIG. 9 shows that when the L-ArgR is reconstituted into a bilayer, the same properties are seen. This demonstrates that a receptor like the L-ArgR is more directly and more readily studied when in the bilayer than when in the native state where its activity must be inferred from secondary neural recordings.
[0139] Reconstitution of the ENaC Protein into Liposomes.
[0140] An amount (0.01 to 0.5 μg) of the sodium channel of interest, including ENaCα, ENaCδ, or specific ratio mixtures derived from the ENaC subunits, α, β, γ, δ, dissolved in 100 mM NaCl/50 mM Tris, pH=7.1 containing one of several detergents such as Tween, CHAPS, Triton, Sodium Cholate, or/and Octyl-glucoside will be added to the liposomes. The ratio of protein to lipid (mass:mass) for measuring channel activity as single cannel events is 1:2000-5000. The ratio for measuring macroscopic properties is in the range of 1:50-100. The protein-lipid mixture will be incubated for 20-30 mins This procedure will be followed because the ENaC protein, being a membrane channel, needs to remain in solution while it is reconstituted into liposomes.
[0141] Reconstituted detergent-free proteo-liposomes containing one or more of the ENaCs can be prepared at least two ways. In one method, they can be formed through centrifugation of the protein:lipid mixture through gel filtration columns. These gel columns are prepared from Sephadex G-50 (fine), swollen overnight, and poured into 5-ml disposable columns (1.5-ml bed volume). Columns are pre-spun in a centrifuge at ˜1,000×g. The protein:lipid mixture is loaded on the top of the column, and proteo-liposomes free of detergent can be recovered by spinning the columns at 700 g for 1 min. Alternatively, detergent can be removed by dialysis. For dialysis, the protein:lipid mixture is loaded into a cassette dialysis unit and the mixture dialyzed overnight against 2000 ml of a Tris/NaCl/sucrose (detergent-free) buffer at 4° C. Phospholipid vesicles containing the protein are expected to form spontaneously as the concentration of detergent decreases during the dialysis.
[0142] Reconstitution of the Channel Proteins from Proteo-Liposomes into a Planar Lipid Bilayer.
[0143] The planar lipid bilayer is formed on an aperture between two aqueous compartments, which for operational purposes are called cis and trans compartments. The voltage generator will be connected to the cis compartment, with an Ag/AgCl electrode, to control membrane potential. The trans compartment (virtual ground) will be connected to the input of the current-measuring amplifier through a second Ag/AgCl electrode.
[0144] Forming the Bilayer.
[0145] A 4:1 mixture of DOPE:DOPC will be dissolved in 25 μl of n-decane (concentration ranging from 15 to 25 mg/ml). This mixture will be kept at room temperature and prepared each day that the experiment is performed. Electrode compartments will be filled with 3 M KCl and the Ag/AgCl electrodes will be placed in the compartment. The cis and trans compartments will be filled with the recording bath solution (100 mM NaCl, 10 Tris, pH 7) and agar bridges will be placed between them and the electrode compartments. To form the bilayer, a droplet of the lipid mixture will be spread onto the hole from the cis side.
[0146] The lipids can be determined to be completely formed around the hole when the resistance increased and the signal is not saturating. To verify that an organized bilayer has formed, the voltage pulses across the bilayer can be applied and "capacitive currents" can be measured. For a hole of 100 μm, the capacitance is expected to be of the order of 50-100 pF. The electrical resistance of the bilayer is expected to be higher than 109 Ohm.
[0147] Reconstitution.
[0148] After the bilayer has formed, 10-15 μl of the proteo-liposomes will be added to the cis side of the bilayer under constant stirring. When channel subunits are incorporated into the bilayer, the currents are expected to change in steps. Macroscopic current will be measured when many channels are incorporated.
[0149] Liposome fusion with the bilayer happens spontaneously, and currents will be able to be recorded within about 5 to 30 minutes. In some cases, it may be necessary to facilitate the liposome fusion by: creating a concentration gradient across the liposome by adding the liposomes formed previously in 300 mM NaCl to a bilayer bathing solution containing 100 mM NaCl, or by creating a concentration gradient across the lipid bilayer by adding 100 mM NaCl to the cis side and 10 mM NaCl to the trans side, or by changing bilayer and/or liposome lipid composition by the addition of negatively charged lipid such as DOPS to the bilayer.
Example 4
Expression of Varying Ratios of ENaC Receptor Subdomains in Lipid Bilayers
[0150] This is a prophetic example. The ENaC is a heteromultimeric complex generally comprised of three subunits: either of subunits α, β, and γ (in most tissues as α2 βγ complex) or subunits δ, β, and γ. These subunits can assemble in varying ratios, often dictated by the tissue source. Varying the relative ratios of the subunits confers unique kinetics and pharmacology upon these channels. Without intending to be limited to any particular theory or mechanism of action, it is believed that the δ subunit replaces the α subunit in many tissues, and that such a substitution may modify particularly the pharmacology of the channel.
[0151] Single Cell Quantitative PCR with Specific Reference to Estimation of the Ratios Of ENaC Subunits.
[0152] While there is no guarantee of a one-to-one correspondence between amount of message and amount of protein, Q-PCR is one tool available for estimation of ENaC ratio. Assuming a salty taste cell is active, it is likely to have a number of copies of a particular subunit. It is likely that the ratio of message copies will be at least approximately that of the protein products. Quantitative single cell PCR can be used to gain a semi-quantitative picture of the relative abundance of message for any proteins of interest. The procedure, although theoretically straight forward, presents a number of challenging obstacles. With taste cells, for example, RNA quality can be problematic because the time-consuming procedure currently used to obtain isolated cells (see above) is conducive to destruction of RNA. To be confident in the experimental technique, the following procedure can be carried out: (1) Design several unique primer pairs for each gene of interest, using only those that have almost identical efficiency under the same PCR conditions for every gene of interest. (2) Construct a primer set (mixture of primer pairs) from the appropriate pairs above that registers as a blank when used in a water control PCR reaction. (3) Collect individual cells (as above) into an RNase-free environment, lyse the cell and reverse transcribe the single cell mRNA content using a commercially available kit. (4) Run a limited (10-25) number of cycles of the first stage PCR with the primer set and condition above, so that all of the reactions are in a linear amplification phase. (5) Dilute the above reaction (100×-1000×), and use an aliquot as template along with a single pair of primers from the set above and perform the second stage of PCR (in duplicate/triplicate) using a Q-PCR machine. (6) A relative quantification method is used for data analysis. Normalization is based on amplification of a genomic DNA that is not translated/transcribed of which there is, by definition, one copy of the gene. The differences in gene expression can be determined by comparing ratio (ΔCt between target gene and genomic reference sequence in sample) differences (ΔΔCts, the differences of ΔCts between two samples).
[0153] A taste bud cell containing message for human ENaC subunits δ, β, and γ, but no message for α was apparent. The Q-PCR trace of this analysis is shown in FIG. 7. From this single cell, it can be concluded that the ENAC of that cell is of multimeric structure, δ1 β1 γ1. However, as these traces are not normalized, the definitive structure may have a different stoichiometric ratio. The best evidence, however, suggests that the human salty taste receptor is composed of δ1 β1 γ1.
[0154] Once sequence confirmation is obtained, the recombinant plasmid can be used as the substrate in a process known as in vitro protein expression (IVPE). This procedure, be it either cell driven or a cell-free system, allows generation of large amounts (mM) of desired protein, in this case, ENaC subunits, δ, α, β, and γ. A Western blot can be used to confirm the identity of the manufactured protein. Analysis of the reaction mixture using an antibody to the protein (a Western blot) is used to confirm that the desired protein has indeed been obtained.
[0155] The desired protein can be isolated and purified. Purification of the protein by affinity chromatography involves chemically linking an antibody to the protein with a column matrix such as Sephadex. Passing the IVPE reaction mixture through the column results in binding of the protein to its antibody on the column. Elution of the column with an appropriate reagent yields the enriched protein. The protein eluate can be quantified by measuring total nitrogen, as in the Kjeldahl procedure. This measure of total nitrogen content is then compared to the protein's absorption at 280 nm to calculate an absorption coefficient. From this point, absorption at 280 nm becomes a convenient and accurate measure of protein concentration.
[0156] Knowing the actual concentration of each subunit of the ENaC allows the combination of these subunits in specific ratios, these having been estimated by the Q-PCR of single cells. As these proteins are membrane associated, they will require some amount of detergent to remain soluble. While their being soluble is required for combining them in specific ratios, detergent will destroy the lipid bilayer into which they need to be reconstituted to measure activity. Thus, reconstitution of the lipid bilayer with the isolated proteins requires that any detergent be removed. Detergent can be removed by any means suitable in the art, such as dialysis as described herein. Reconstitution of isolated proteins into lipid membranes has been described (Grosvenor, W et al. (2004) BMC Neurosci. 5:2202-5), and summarized in the examples above. Because the subunits for human ENaC are synthesized, an advantage is gained as careful control over the composition an ratios of any putative salt taste receptor subunits can be exerted.
[0157] The present invention is not limited to the embodiments described and exemplified above, but is capable of variation and modification within the scope of the appended claims.
TABLE-US-00002 Sequence Listing SEQ ID NO: 1 nt for human alpha atggagggga acaagctgga ggagcaggac tctagccctc cacagtccac tccagggctc atgaagggga acaagcgtga ggagcagggg ctgggccccg aacctgcggc gccccagcag cccacggcgg aggaggaggc cctgatcgag ttccaccgct cctaccgaga gctcttcgag ttcttctgca acaacaccac catccacggc gccatccgcc tggtgtgctc ccagcacaac cgcatgaaga cggccttctg ggcagtgctg tggctctgca cctttggcat gatgtactgg caattcggcc tgcttttcgg agagtacttc agctaccccg tcagcctcaa catcaacctc aactcggaca agctcgtctt ccccgcagtg accatctgca ccctcaatcc ctacaggtac ccggaaatta aagaggagct ggaggagctg gaccgcatca cagagcagac gctctttgac ctgtacaaat acagctcctt caccactctc gtggccggct cccgcagccg tcgcgacctg cgggggactc tgccgcaccc cttgcagcgc ctgagggtcc cgcccccgcc tcacggggcc cgtcgagccc gtagcgtggc ctccagcttg cgggacaaca acccccaggt ggactggaag gactggaaga tcggcttcca gctgtgcaac cagaacaaat cggactgctt ctaccagaca tactcatcag gggtggatgc ggtgagggag tggtaccgct tccactacat caacatcctg tcgaggctgc cagagactct gccatccctg gaggaggaca cgctgggcaa cttcatcttc gcctgccgct tcaaccaggt ctcctgcaac caggcgaatt actctcactt ccaccacccg atgtatggaa actgctatac tttcaatgac aagaacaact ccaacctctg gatgtcttcc atgcctggaa tcaacaacgg tctgtccctg atgctgcgcg cagagcagaa tgacttcatt cccctgctgt ccacagtgac tggggcccgg gtaatggtgc acgggcagga tgaacctgcc tttatggatg atggtggctt taacttgcgg cctggcgtgg agacctccat cagcatgagg aaggaaaccc tggacagact tgggggcgat tatggcgact gcaccaagaa tggcagtgat gttcctgttg agaaccttta cccttcaaag tacacacagc aggtgtgtat tcactcctgc ttccaggaga gcatgatcaa ggagtgtggc tgtgcctaca tcttctatcc gcggccccag aacgtggagt actgtgacta cagaaagcac agttcctggg ggtactgcta ctataagctc caggttgact tctcctcaga ccacctgggc tgtttcacca agtgccggaa gccatgcagc gtgaccagct accagctctc tgctggttac tcacgatggc cctcggtgac atcccaggaa tgggtcttcc agatgctatc gcgacagaac aattacaccg tcaacaacaa gagaaatgga gtggccaaag tcaacatctt cttcaaggag ctgaactaca aaaccaattc tgagtctccc tctgtcacga tggtcaccct cctgtccaac ctgggcagcc agtggagcct gtggttcggc tcctcggtgt tgtctgtggt ggagatggct gagctcgtct ttgacctgct ggtcatcatg ttcctcatgc tgctccgaag gttccgaagc cgatactggt ctccaggccg agggggcagg ggtgctcagg aggtagcctc caccctggca tcctcccctc cttcccactt ctgcccccac cccatgtctc tgtccttgtc ccagccaggc cctgctccct ctccagcctt gacagcccct ccccctgcct atgccaccct gggcccccgc ccatctccag ggggctctgc aggggccagt tcctccacct gtcctctggg ggggccctga SEQ ID NO: 2 pro for human alpha MEGNKLEEQD SSPPQSTPGL MKGNKREEQG LGPEPAAPQQ PTAEEEALIE FHRSYRELFE FFCNNTTIHG AIRLVCSQHN RMKTAFWAVL WLCTFGMMYW QFGLLFGEYF SYPVSLNINL NSDKLVFPAV TICTLNPYRY PEIKEELEEL DRITEQTLFD LYKYSSFTTL VAGSRSRRDL RGTLPHPLQR LRVPPPPHGA RRARSVASSL RDNNPQVDWK DWKIGFQLCN QNKSDCFYQT YSSGVDAVRE WYRFHYINIL SRLPETLPSL EEDTLGNFIF ACRFNQVSCN QANYSHFHHP MYGNCYTFND KNNSNLWMSS MPGINNGLSL MLRAEQNDFI PLLSTVTGAR VMVHGQDEPA FMDDGGFNLR PGVETSISMR KETLDRLGGD YGDCTKNGSD VPVENLYPSK YTQQVCIHSC FQESMIKECG CAYIFYPRPQ NVEYCDYRKH SSWGYCYYKL QVDFSSDHLG CFTKCRKPCS VTSYQLSAGY SRWPSVTSQE WVFQMLSRQN NYTVNNKRNG VAKVNIFFKE LNYKTNSESP SVTMVTLLSN LGSQWSLWFG SSVLSVVEMA ELVFDLLVIM FLMLLRRFRS RWPSPGRGGR GAQEVASTLA SSPPSHFCPH PMSLSLSQPG PAPSPALTAP PPAYATLGPR PSPGGSAGAS SSTCPLGGP SEQ ID NO: 3 nt for human beta atgcacgtga agaagtacct gctgaagggc ctgcatcggc tgcagaaggg ccccggctac acgtacaagg agctgctggt gtggtactgc gacaacacca acacccacgg ccccaagcgc atcatctgtg aggggcccaa gaagaaagcc atgtggttcc tgctcaccct gctcttcgcc gccctcgtct gctggcagtg gggcatcttc atcaggacct acttgagctg ggaggtcagc gtctccctct ccgtaggctt caagaccatg gacttccccg ccgtcaccat ctgcaatgct agccccttca agtattccaa aatcaagcat ttgctgaagg acctggatga gctgatggaa gctgtcctgg agagaatcct ggctcctgag ctaagccatg ccaatgccac caggaacctg aacttctcca tctggaacca cacacccctg gtccttattg atgaacggaa cccccaccac cccatggtcc ttgatctctt tggagacaac cacaatggct taacaagcag ctcagcatca gaaaagatct gtaatgccca cgggtgcaaa atggccatga gactatgtag cctcaacagg acccagtgta ccttccggaa cttcaccagt gctacccagg cattgacaga gtggtacatc ctgcaggcca ccaacatctt tgcacaggtg ccacagcagg agctagtaga gatgagctac cccggcgagc agatgatcct ggcctgccta ttcggagctg agccctgcaa ctaccggaac ttcacgtcca tcttctaccc tcactatggc aactgttaca tcttcaactg gggcatgaca gagaaggcac ttccttcggc caaccctgga actgaattcg gcctgaagtt gatcctggac ataggccagg aagactacgt ccccttcctt gcgtccacgg ccggggtcag gctgatgctt cacgagcaga ggtcataccc cttcatcaga gatgagggca tctacGccat gtcggggaca gagacgtcca tcggggtact cgtggacaag cttcagcgca tgggggagcc ctacagcccg tgcaccgtga atggttctga ggtccccgtc caaaacttct acagtgacta caacacgacc tactccatcc aggcctgtct tcgctcctgc ttccaagacc acatgatccg taactgcaac tgtggccact acctgtaccc actGccccgt ggggagaaat actgcaacaa ccgggacttc ccagactggg cccattgcta ctcagatcta cagatgagcg tggcgcagag agagacctgc attggcatgt gcaaggagtc ctgcaatgac acccagtaca agatgaccat ctccatggct gactggcctt ctgaggcctc cgaggactgg attttccacg tcttgtctca ggagcgggac caaagcacca atatcaccct gagcaggaag ggaattgtca agctcaacat ctacttccaa gaatttaact atcgcaccat tgaagaatca gcagccaata acatcgtctg gctgctctcg aatctgggtg gccagtttgg cttctggatg gggggctctg tgctgtgcct catcgagttt ggggagatca tcatcgactt tgtgtggatc accatcatca agctggtggc cttggccaag agcctacggc agcggcgagc ccaagccagc tacgctggcc caccgcccac cgtggccgag ctggtggagg cccacaccaa ctttggcttc cagcctgaca cggccccccg cagccccaac actgggccct accccagtga gcaggccctg cccatcccag gcaccccgcc ccccaactat gactccctgc gtctgcagcc gctggacgtc atcgagtctg acagtgaggg tgatgccatc taa SEQ ID NO: 4 pro for human beta MHVKKYLLKG LHRLQKGPGY TYKELLVWYC DNTNTHGPKR IICEGPKKKA MWFLLTLLFA ALVCWQWGIF IRTYLSWEVS VSLSVGFKTM DFPAVTICNA SPFKYSKIKH LLKDLDELME AVLERILAPE LSHANATRNL NFSIWNHTPL VLIDERNPHH PMVLDLFGDN HNGLTSSSAS EKICNAHGCK MAMRLCSLNR TQCTFRNFTS ATQALTEWYI LQATNIFAQV PQQELVEMSY PGEQMILACL FGAEPCNYRN FTSIFYPHYG NCYIFNWGMT EKALPSANPG TEFGLKLILD IGQEDYVPFL ASTAGVRLML HEQRSYPFIR DEGIYAMSGT ETSIGVLVDK LQRMGEPYSP CTVNGSEVPV QNFYSDYNTT YSIQACLRSC FQDHMIRNCN CGHYLYPLPR GEKYCNNRDF PDWAHCYSDL QMSVAQRETC IGMCKESCND TQYKMTISMA DWPSEASEDW IFHVLSQERD QSTNITLSRK GIVKLNIYFQ EFNYRTIEES AANNIVWLLS NLGGQFGFWM GGSVLCLIEF GEIIIDFVWI TIIKLVALAK SLRQRRAQAS YAGPPPTVAE LVEAHTNFGF QPDTAPRSPN TGPYPSEQAL PIPGTPPPNY DSLRLQPLDV IESDSEGDAI SEQ ID NO: 5 nt for human taste gamma atggcacccg gagagaagat caaagccaaa atcaagaaga atctgcccgt gacgggccct caggcgccga ccattaaaga gctgatgcgg tggtactgcc tcaacaccaa cacccatggc tgtcgccgca tcgtggtgtc ccgcggccgt ctgcgccgcc tcctctggat cgggttcaca ctgactgccg tggccctcat cctctggcag tgcgccctcc tcgtcttctc cttctatact gtctcagttt ccatcaaagt ccacttccgg aagctggatt ttcctgcagt caccatctgc aacatcaacc cctacaagta cagcaccgtt cgccaccttc tagctgactt ggaacaggag accagagagg ccctgaagtc cctgtatggc tttccagagt cccggaagcg ccgagaggcg gagtcctgga actccgtctc agagggaaag cagcctagat tctcccaccg gattccgctg ctgatctttg atcaggatga gaagggcaag gccagggact tcttcacagg gaggaagcgg aaagtcggcg gtagcatcat tcacaaggct tcaaatgtca tgcacatcga gtccaagcaa gtggtgggat tccaactgtg ctcaaatgac acctccgact gtgccaccta caccttcagc tcgggaatca atgccattca ggagtggtat aagctacact acatgaacat catggcacag gtgcctctgg agaagaaaat caacatgagc tattctgctg aggagctgct ggtgacctgc ttctttgatg gagtgtcctg tgatgccagg aatttcacgc ttttccacca cccgatgcat gggaattgct atactttcaa caacagagaa aatgagacca ttctcagcac ctccatgggg ggcagcgaat atgggctgca agtcattttg tacataaacg aagaggaata caacccattc ctcgtgtcct ccactggagc taaggtgatc atccatcggc aggatgagta tcccttcgtc gaagatgtgg gaacagagat tgagacagca atggtcacct ctataggaat gcacctgaca gagtccttca agctgagtga gccctacagt cagtgcacgg aggacgggag tgacgtgcca atcaggaaca tctacaacgc tgcctactcg ctccagatct gccttcattc atgcttccag acaaagatgg tggagaaatg tgggtgtgcc cagtacagcc agcctctacc tcctgcagcc aactactgca actaccagca gcaccccaac tggatgtatt gttactacca actgcatcga gcctttgtcc aggaagagct gggctgccag tctgtgtgca aggaagcctg cagctttaaa gagtggacac taaccacaag cctggcacaa tggccatctg tggtttcgga gaagtggttg ctgcctgttc tcacttggga ccaaggccgg caagtaaaca aaaagctcaa caagacagac ttggccaaac tcttgatatt ctacaaagac ctgaaccaga gatccatcat ggagagccca gccaacagta ttgagatgct tctgtccaac ttcggtggcc agctgggcct gtggatgagc tgctctgttg tctgcgtcat cgagatcatc gaggtcttct tcattgactt cttctctatc attgcccgcc gccagtggca gaaagccaag gagtggtggg cctggaaaca ggctccccca tgtccagaag ctccccgtag cccacagggc caggacaatc cagccctgga tatagacgat gacctaccca ctttcaactc tgctttgcac ctgcctccag ccctaggaac ccaagtgccc ggcacaccgc cccccaaata caataccttg cgcttggaga gggccttttc caaccagctc acagataccc agatgctaga tgagctctga SEQ ID NO: 6 pro for human taste gamma MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 7 nt for human kidney delta atggctgagc accgaagcat ggacgggaga atggaagcag ccacacgggg gggctctcac ctccaggctg cagcccagac gccccccagg ccggggccac catcagcacc accaccacca cccaaggagg ggcaccagga ggggctggtg gagctgcccg cctcgttccg ggagctgctc accttcttct gcaccaatgc caccatccac ggcgccatcc gcctggtctg ctcccgcggg aaccgcctca agacgacgtc ctgggggctg ctgtccctgg gagccctggt cgcgctctgc tggcagctgg ggctcctctt tgagcgtcac tggcaccgcc
cggtcctcat ggccgtctct gtgcactcgg agcgcaagct gctcccgctg gtcaccctgt gtgacgggaa cccacgtcgg ccgagtccgg tcctccgcca tctggagctg ctggacgagt ttgccaggga gaacattgac tccctgtaca acgtcaacct cagcaaaggc agagccgccc tctccgccac tgtcccccgc cacgagcccc ccttccacct ggaccgggag atccgtctgc agaggctgag ccactcgggc agccgggtca gagtggggtt cagactgtgc aacagcacgg gcggcgactg cttttaccga ggctacacgt caggcgtggc ggctgtccag gactggtacc acttccacta tgtggatatc ctggccctgc tgcccgcggc atgggaggac agccacggga gccaggacgg ccacttcgtc ctctcctgca gttacgatgg cctggactgc caggcccgac agttccggac cttccaccac cccacctacg gcagctgcta cacggtcgat ggcgtctgga cagctcagcg ccccggcatc acccacggag tcggcctggt cctcagggtt gagcagcagc ctcacctccc tctgctgtcc acgctggccg gcatcagggt catggttcac ggccgtaacc acacgccctt cctggggcac cacagcttca gcgtccggcc agggacggag gccaccatca gcatccgaga ggacgaggtg caccggctcg ggagccccta cggccactgc accgccggcg gggaaggcgt ggaggtggag ctgctacaca acacctccta caccaggcag gcctgcctgg tgtcctgctt ccagcagctg atggtggaga cctgctcctg tggctactac ctccaccctc tgccggcggg ggctgagtac tgcagctctg cccggcaccc tgcctgggga cactgcttct accgcctcta ccaggacctg gagacccacc ggctcccctg tacctcccgc tgccccaggc cctgcaggga gtctgcattc aagctctcca ctgggacctc caggtggcct tccgccaagt cagctggatg gactctggcc acgctaggtg aacaggggct gccgcatcag agccacagac agaggagcag cctggccaaa atcaacatcg tctaccagga gctcaactac cgctcagtgg aggaggcgcc cgtgtactcg gtgccgcagc tgctctccgc catgggcagc ctctacagcc tgtggtttgg ggcctccgtc ctctccctcc tggagctcct ggagctgctg ctcgatgctt ctgccctcac cctggtgcta ggcggccgcc ggctccgcag ggcgtggttc tcctggccca gagccagccc tgcctcaggg gcgtccagca tcaagccaga ggccagtcag atgcccccgc ctgcaggcgg cacgtcagat gacccggagc ccagcgggcc tcatctccca cgggtgatgc ttccaggggt tctggcggga gtctcagccg aagagagctg ggctgggccc cagccccttg agactctgga cacctga Note: tac->tgc provides Tyr->Cys at 532 SEQ ID NO: 8 pro for human kidney delta (with Y532) MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LYSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 9 pro for humanbrain delta (with C532) MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 10 motif for human kidney alpha MGSQWSLWFGA SEQ ID NO: 11 motif for human kidney delta MGSLYSLWFGA SEQ ID NO: 12 motif for human taste bud delta MGSLCSLWFGA SEQ ID NO: 13 pro for human taste gamma (Gamma A) MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPP LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS LGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSLK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVVEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 14 pro for human taste gamma (Gamma B) MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYA VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNSL TDTQMLDEL SEQ ID NO: 15 pro for human taste gamma (Gamma C) MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD GLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 16 pro for human lung gamma (X87160) MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPSV EDVGTEIETT MVTSIGMHLT ESFKLSEPSS QCTEGGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACRFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 17 pro for human delta subject DENACA MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTIHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATIRIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLHRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 18 pro for human delta subject DENACD MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 19 pro for human delta subject DENAE MAEHRSMDGR MEAATRGGPH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPP HEPPFHLDRE IRLQSLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDK SEQ ID NO: 20 pro for human delta subject DENACG MAEHRSMDGR MEAATRGGPH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR
PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQGLNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 21 pro for human delta subject DENACH MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GTIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPP HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVQ LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYRDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEAGQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 22 pro for human delta subject DENACI MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ GHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 23 pro for human delta subject DENACJ MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPP HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVQ PLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 24 pro for human delta subject DENACT MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYSVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ PCLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 25 pro for human delta subject DENACV MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTFHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATISIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CGSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLRRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 26 pro for human delta subject DENACW MAEHRSMDGR MEAATRGGSH LQAAAQTPPR PGPPSAPPPP PKEGHQEGLV ELPASFRELL TFFCTNATIH GAIRLVCSRG NRLKTTSWGL LSLGALVALC WQLGLLFERH WHRPVLMAVS VHSERKLLPL VTLCDGNPRR PSPVLRHLEL LDEFARENID SLYNVNLSKG RAALSATVPR HEPPFHLDRE IRLQRLSHSG SRVRVGFRLC NSTGGDCFYR GYTSGVAAVQ DWYHFHYVDI LALLPAAWED SHGSQDGHFV LSCSYDGLDC QARQFRTIHH PTYGSCYTVD GVWTAQRPGI THGVGLVLRV EQQPHLPLLS TLAGIRVMVH GRNHTPFLGH HSFSVRPGTE ATIRIREDEV HRLGSPYGHC TAGGEGVEVE LLHNTSYTRQ ACLVSCFQQL MVETCSCGYY LHPLPAGAEY CSSARHPAWG HCFYRLYQDL ETHRLPCTSR CPRPCRESAF KLSTGTSRWP SAKSAGWTLA TLGEQGLPHQ SHRQRSSLAK INIVYQELNY RSVEEAPVYS VPQLLSAMGS LCSLWFGASV LSLLELLELL LDASALTLVL GGRRLHRAWF SWPRASPASG ASSIKPEASQ MPPPAGGTSD DPEPSGPHLP RVMLPGVLAG VSAEESWAGP QPLETLDT SEQ ID NO: 27 pro for human gamma subject GENACA MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 28 pro for human gamma subject GENACB MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVNFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 29 pro for human gamma subject GENACD MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 30 pro for human gamma subject GENACE MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGE ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP STPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 31 pro for human gamma subject GENACG MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYA VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI TARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNSL TDTQMLDEL SEQ ID NO: 32 pro for human gamma subject GENACH MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG
FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD GLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 33 pro for human gamma subject GENACJ MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPP LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS LGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSLK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI TARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 34 pro for human gamma subject GENACT MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 35 pro for human gamma subject GENACV MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILWQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL SEQ ID NO: 36 pro for human gamma subject GENACW MAPGEKIKAK IKKNLPVTGP QAPTIKELMR WYCLNTNTHG CRRIVVSRGR LRRLLWIGFT LTAVALILRQ CALLVFSFYT VSVSIKVHFR KLDFPAVTIC NINPYKYSTV RHLLADLEQE TREALKSLYG FPESRKRREA ESWNSVSEGK QPRFSHRIPL LIFDQDEKGK ARDFFTGRKR KVGGSIIHKA SNVMHIESKQ VVGFQLCSND TSDCATYTFS SGINAIQEWY KLHYMNIMAQ VPLEKKINMS YSAEELLVTC FFDGVSCDAR NFTLFHHPMH GNCYTFNNRE NETILSTSMG GSEYGLQVIL YINEEEYNPF LVSSTGAKVI IHRQDEYPFV EDVGTEIETA MVTSIGMHLT ESFKLSEPYS QCTEDGSDVP IRNIYNAAYS LQICLHSCFQ TKMVEKCGCA QYSQPLPPAA NYCNYQQHPN WMYCYYQLHR AFVQEELGCQ SVCKEACSFK EWTLTTSLAQ WPSVVSEKWL LPVLTWDQGR QVNKKLNKTD LAKLLIFYKD LNQRSIMESP ANSIEMLLSN FGGQLGLWMS CSVVCVIEII EVFFIDFFSI IARRQWQKAK EWWAWKQAPP CPEAPRSPQG QDNPALDIDD DLPTFNSALH LPPALGTQVP GTPPPKYNTL RLERAFSNQL TDTQMLDEL
Sequence CWU
1
1
3812010DNAHomo sapiens 1atggagggga acaagctgga ggagcaggac tctagccctc
cacagtccac tccagggctc 60atgaagggga acaagcgtga ggagcagggg ctgggccccg
aacctgcggc gccccagcag 120cccacggcgg aggaggaggc cctgatcgag ttccaccgct
cctaccgaga gctcttcgag 180ttcttctgca acaacaccac catccacggc gccatccgcc
tggtgtgctc ccagcacaac 240cgcatgaaga cggccttctg ggcagtgctg tggctctgca
cctttggcat gatgtactgg 300caattcggcc tgcttttcgg agagtacttc agctaccccg
tcagcctcaa catcaacctc 360aactcggaca agctcgtctt ccccgcagtg accatctgca
ccctcaatcc ctacaggtac 420ccggaaatta aagaggagct ggaggagctg gaccgcatca
cagagcagac gctctttgac 480ctgtacaaat acagctcctt caccactctc gtggccggct
cccgcagccg tcgcgacctg 540cgggggactc tgccgcaccc cttgcagcgc ctgagggtcc
cgcccccgcc tcacggggcc 600cgtcgagccc gtagcgtggc ctccagcttg cgggacaaca
acccccaggt ggactggaag 660gactggaaga tcggcttcca gctgtgcaac cagaacaaat
cggactgctt ctaccagaca 720tactcatcag gggtggatgc ggtgagggag tggtaccgct
tccactacat caacatcctg 780tcgaggctgc cagagactct gccatccctg gaggaggaca
cgctgggcaa cttcatcttc 840gcctgccgct tcaaccaggt ctcctgcaac caggcgaatt
actctcactt ccaccacccg 900atgtatggaa actgctatac tttcaatgac aagaacaact
ccaacctctg gatgtcttcc 960atgcctggaa tcaacaacgg tctgtccctg atgctgcgcg
cagagcagaa tgacttcatt 1020cccctgctgt ccacagtgac tggggcccgg gtaatggtgc
acgggcagga tgaacctgcc 1080tttatggatg atggtggctt taacttgcgg cctggcgtgg
agacctccat cagcatgagg 1140aaggaaaccc tggacagact tgggggcgat tatggcgact
gcaccaagaa tggcagtgat 1200gttcctgttg agaaccttta cccttcaaag tacacacagc
aggtgtgtat tcactcctgc 1260ttccaggaga gcatgatcaa ggagtgtggc tgtgcctaca
tcttctatcc gcggccccag 1320aacgtggagt actgtgacta cagaaagcac agttcctggg
ggtactgcta ctataagctc 1380caggttgact tctcctcaga ccacctgggc tgtttcacca
agtgccggaa gccatgcagc 1440gtgaccagct accagctctc tgctggttac tcacgatggc
cctcggtgac atcccaggaa 1500tgggtcttcc agatgctatc gcgacagaac aattacaccg
tcaacaacaa gagaaatgga 1560gtggccaaag tcaacatctt cttcaaggag ctgaactaca
aaaccaattc tgagtctccc 1620tctgtcacga tggtcaccct cctgtccaac ctgggcagcc
agtggagcct gtggttcggc 1680tcctcggtgt tgtctgtggt ggagatggct gagctcgtct
ttgacctgct ggtcatcatg 1740ttcctcatgc tgctccgaag gttccgaagc cgatactggt
ctccaggccg agggggcagg 1800ggtgctcagg aggtagcctc caccctggca tcctcccctc
cttcccactt ctgcccccac 1860cccatgtctc tgtccttgtc ccagccaggc cctgctccct
ctccagcctt gacagcccct 1920ccccctgcct atgccaccct gggcccccgc ccatctccag
ggggctctgc aggggccagt 1980tcctccacct gtcctctggg ggggccctga
20102669PRTHomo Sapiens 2Met Glu Gly Asn Lys Leu
Glu Glu Gln Asp Ser Ser Pro Pro Gln Ser 1 5
10 15 Thr Pro Gly Leu Met Lys Gly Asn Lys Arg Glu
Glu Gln Gly Leu Gly 20 25
30 Pro Glu Pro Ala Ala Pro Gln Gln Pro Thr Ala Glu Glu Glu Ala
Leu 35 40 45 Ile
Glu Phe His Arg Ser Tyr Arg Glu Leu Phe Glu Phe Phe Cys Asn 50
55 60 Asn Thr Thr Ile His Gly
Ala Ile Arg Leu Val Cys Ser Gln His Asn 65 70
75 80 Arg Met Lys Thr Ala Phe Trp Ala Val Leu Trp
Leu Cys Thr Phe Gly 85 90
95 Met Met Tyr Trp Gln Phe Gly Leu Leu Phe Gly Glu Tyr Phe Ser Tyr
100 105 110 Pro Val
Ser Leu Asn Ile Asn Leu Asn Ser Asp Lys Leu Val Phe Pro 115
120 125 Ala Val Thr Ile Cys Thr Leu
Asn Pro Tyr Arg Tyr Pro Glu Ile Lys 130 135
140 Glu Glu Leu Glu Glu Leu Asp Arg Ile Thr Glu Gln
Thr Leu Phe Asp 145 150 155
160 Leu Tyr Lys Tyr Ser Ser Phe Thr Thr Leu Val Ala Gly Ser Arg Ser
165 170 175 Arg Arg Asp
Leu Arg Gly Thr Leu Pro His Pro Leu Gln Arg Leu Arg 180
185 190 Val Pro Pro Pro Pro His Gly Ala
Arg Arg Ala Arg Ser Val Ala Ser 195 200
205 Ser Leu Arg Asp Asn Asn Pro Gln Val Asp Trp Lys Asp
Trp Lys Ile 210 215 220
Gly Phe Gln Leu Cys Asn Gln Asn Lys Ser Asp Cys Phe Tyr Gln Thr 225
230 235 240 Tyr Ser Ser Gly
Val Asp Ala Val Arg Glu Trp Tyr Arg Phe His Tyr 245
250 255 Ile Asn Ile Leu Ser Arg Leu Pro Glu
Thr Leu Pro Ser Leu Glu Glu 260 265
270 Asp Thr Leu Gly Asn Phe Ile Phe Ala Cys Arg Phe Asn Gln
Val Ser 275 280 285
Cys Asn Gln Ala Asn Tyr Ser His Phe His His Pro Met Tyr Gly Asn 290
295 300 Cys Tyr Thr Phe Asn
Asp Lys Asn Asn Ser Asn Leu Trp Met Ser Ser 305 310
315 320 Met Pro Gly Ile Asn Asn Gly Leu Ser Leu
Met Leu Arg Ala Glu Gln 325 330
335 Asn Asp Phe Ile Pro Leu Leu Ser Thr Val Thr Gly Ala Arg Val
Met 340 345 350 Val
His Gly Gln Asp Glu Pro Ala Phe Met Asp Asp Gly Gly Phe Asn 355
360 365 Leu Arg Pro Gly Val Glu
Thr Ser Ile Ser Met Arg Lys Glu Thr Leu 370 375
380 Asp Arg Leu Gly Gly Asp Tyr Gly Asp Cys Thr
Lys Asn Gly Ser Asp 385 390 395
400 Val Pro Val Glu Asn Leu Tyr Pro Ser Lys Tyr Thr Gln Gln Val Cys
405 410 415 Ile His
Ser Cys Phe Gln Glu Ser Met Ile Lys Glu Cys Gly Cys Ala 420
425 430 Tyr Ile Phe Tyr Pro Arg Pro
Gln Asn Val Glu Tyr Cys Asp Tyr Arg 435 440
445 Lys His Ser Ser Trp Gly Tyr Cys Tyr Tyr Lys Leu
Gln Val Asp Phe 450 455 460
Ser Ser Asp His Leu Gly Cys Phe Thr Lys Cys Arg Lys Pro Cys Ser 465
470 475 480 Val Thr Ser
Tyr Gln Leu Ser Ala Gly Tyr Ser Arg Trp Pro Ser Val 485
490 495 Thr Ser Gln Glu Trp Val Phe Gln
Met Leu Ser Arg Gln Asn Asn Tyr 500 505
510 Thr Val Asn Asn Lys Arg Asn Gly Val Ala Lys Val Asn
Ile Phe Phe 515 520 525
Lys Glu Leu Asn Tyr Lys Thr Asn Ser Glu Ser Pro Ser Val Thr Met 530
535 540 Val Thr Leu Leu
Ser Asn Leu Gly Ser Gln Trp Ser Leu Trp Phe Gly 545 550
555 560 Ser Ser Val Leu Ser Val Val Glu Met
Ala Glu Leu Val Phe Asp Leu 565 570
575 Leu Val Ile Met Phe Leu Met Leu Leu Arg Arg Phe Arg Ser
Arg Trp 580 585 590
Pro Ser Pro Gly Arg Gly Gly Arg Gly Ala Gln Glu Val Ala Ser Thr
595 600 605 Leu Ala Ser Ser
Pro Pro Ser His Phe Cys Pro His Pro Met Ser Leu 610
615 620 Ser Leu Ser Gln Pro Gly Pro Ala
Pro Ser Pro Ala Leu Thr Ala Pro 625 630
635 640 Pro Pro Ala Tyr Ala Thr Leu Gly Pro Arg Pro Ser
Pro Gly Gly Ser 645 650
655 Ala Gly Ala Ser Ser Ser Thr Cys Pro Leu Gly Gly Pro
660 665 31923DNAHomo Sapiens 3atgcacgtga
agaagtacct gctgaagggc ctgcatcggc tgcagaaggg ccccggctac 60acgtacaagg
agctgctggt gtggtactgc gacaacacca acacccacgg ccccaagcgc 120atcatctgtg
aggggcccaa gaagaaagcc atgtggttcc tgctcaccct gctcttcgcc 180gccctcgtct
gctggcagtg gggcatcttc atcaggacct acttgagctg ggaggtcagc 240gtctccctct
ccgtaggctt caagaccatg gacttccccg ccgtcaccat ctgcaatgct 300agccccttca
agtattccaa aatcaagcat ttgctgaagg acctggatga gctgatggaa 360gctgtcctgg
agagaatcct ggctcctgag ctaagccatg ccaatgccac caggaacctg 420aacttctcca
tctggaacca cacacccctg gtccttattg atgaacggaa cccccaccac 480cccatggtcc
ttgatctctt tggagacaac cacaatggct taacaagcag ctcagcatca 540gaaaagatct
gtaatgccca cgggtgcaaa atggccatga gactatgtag cctcaacagg 600acccagtgta
ccttccggaa cttcaccagt gctacccagg cattgacaga gtggtacatc 660ctgcaggcca
ccaacatctt tgcacaggtg ccacagcagg agctagtaga gatgagctac 720cccggcgagc
agatgatcct ggcctgccta ttcggagctg agccctgcaa ctaccggaac 780ttcacgtcca
tcttctaccc tcactatggc aactgttaca tcttcaactg gggcatgaca 840gagaaggcac
ttccttcggc caaccctgga actgaattcg gcctgaagtt gatcctggac 900ataggccagg
aagactacgt ccccttcctt gcgtccacgg ccggggtcag gctgatgctt 960cacgagcaga
ggtcataccc cttcatcaga gatgagggca tctacgccat gtcggggaca 1020gagacgtcca
tcggggtact cgtggacaag cttcagcgca tgggggagcc ctacagcccg 1080tgcaccgtga
atggttctga ggtccccgtc caaaacttct acagtgacta caacacgacc 1140tactccatcc
aggcctgtct tcgctcctgc ttccaagacc acatgatccg taactgcaac 1200tgtggccact
acctgtaccc actgccccgt ggggagaaat actgcaacaa ccgggacttc 1260ccagactggg
cccattgcta ctcagatcta cagatgagcg tggcgcagag agagacctgc 1320attggcatgt
gcaaggagtc ctgcaatgac acccagtaca agatgaccat ctccatggct 1380gactggcctt
ctgaggcctc cgaggactgg attttccacg tcttgtctca ggagcgggac 1440caaagcacca
atatcaccct gagcaggaag ggaattgtca agctcaacat ctacttccaa 1500gaatttaact
atcgcaccat tgaagaatca gcagccaata acatcgtctg gctgctctcg 1560aatctgggtg
gccagtttgg cttctggatg gggggctctg tgctgtgcct catcgagttt 1620ggggagatca
tcatcgactt tgtgtggatc accatcatca agctggtggc cttggccaag 1680agcctacggc
agcggcgagc ccaagccagc tacgctggcc caccgcccac cgtggccgag 1740ctggtggagg
cccacaccaa ctttggcttc cagcctgaca cggccccccg cagccccaac 1800actgggccct
accccagtga gcaggccctg cccatcccag gcaccccgcc ccccaactat 1860gactccctgc
gtctgcagcc gctggacgtc atcgagtctg acagtgaggg tgatgccatc 1920taa
19234640PRTHomo
Sapiens 4Met His Val Lys Lys Tyr Leu Leu Lys Gly Leu His Arg Leu Gln Lys
1 5 10 15 Gly Pro
Gly Tyr Thr Tyr Lys Glu Leu Leu Val Trp Tyr Cys Asp Asn 20
25 30 Thr Asn Thr His Gly Pro Lys
Arg Ile Ile Cys Glu Gly Pro Lys Lys 35 40
45 Lys Ala Met Trp Phe Leu Leu Thr Leu Leu Phe Ala
Ala Leu Val Cys 50 55 60
Trp Gln Trp Gly Ile Phe Ile Arg Thr Tyr Leu Ser Trp Glu Val Ser 65
70 75 80 Val Ser Leu
Ser Val Gly Phe Lys Thr Met Asp Phe Pro Ala Val Thr 85
90 95 Ile Cys Asn Ala Ser Pro Phe Lys
Tyr Ser Lys Ile Lys His Leu Leu 100 105
110 Lys Asp Leu Asp Glu Leu Met Glu Ala Val Leu Glu Arg
Ile Leu Ala 115 120 125
Pro Glu Leu Ser His Ala Asn Ala Thr Arg Asn Leu Asn Phe Ser Ile 130
135 140 Trp Asn His Thr
Pro Leu Val Leu Ile Asp Glu Arg Asn Pro His His 145 150
155 160 Pro Met Val Leu Asp Leu Phe Gly Asp
Asn His Asn Gly Leu Thr Ser 165 170
175 Ser Ser Ala Ser Glu Lys Ile Cys Asn Ala His Gly Cys Lys
Met Ala 180 185 190
Met Arg Leu Cys Ser Leu Asn Arg Thr Gln Cys Thr Phe Arg Asn Phe
195 200 205 Thr Ser Ala Thr
Gln Ala Leu Thr Glu Trp Tyr Ile Leu Gln Ala Thr 210
215 220 Asn Ile Phe Ala Gln Val Pro Gln
Gln Glu Leu Val Glu Met Ser Tyr 225 230
235 240 Pro Gly Glu Gln Met Ile Leu Ala Cys Leu Phe Gly
Ala Glu Pro Cys 245 250
255 Asn Tyr Arg Asn Phe Thr Ser Ile Phe Tyr Pro His Tyr Gly Asn Cys
260 265 270 Tyr Ile Phe
Asn Trp Gly Met Thr Glu Lys Ala Leu Pro Ser Ala Asn 275
280 285 Pro Gly Thr Glu Phe Gly Leu Lys
Leu Ile Leu Asp Ile Gly Gln Glu 290 295
300 Asp Tyr Val Pro Phe Leu Ala Ser Thr Ala Gly Val Arg
Leu Met Leu 305 310 315
320 His Glu Gln Arg Ser Tyr Pro Phe Ile Arg Asp Glu Gly Ile Tyr Ala
325 330 335 Met Ser Gly Thr
Glu Thr Ser Ile Gly Val Leu Val Asp Lys Leu Gln 340
345 350 Arg Met Gly Glu Pro Tyr Ser Pro Cys
Thr Val Asn Gly Ser Glu Val 355 360
365 Pro Val Gln Asn Phe Tyr Ser Asp Tyr Asn Thr Thr Tyr Ser
Ile Gln 370 375 380
Ala Cys Leu Arg Ser Cys Phe Gln Asp His Met Ile Arg Asn Cys Asn 385
390 395 400 Cys Gly His Tyr Leu
Tyr Pro Leu Pro Arg Gly Glu Lys Tyr Cys Asn 405
410 415 Asn Arg Asp Phe Pro Asp Trp Ala His Cys
Tyr Ser Asp Leu Gln Met 420 425
430 Ser Val Ala Gln Arg Glu Thr Cys Ile Gly Met Cys Lys Glu Ser
Cys 435 440 445 Asn
Asp Thr Gln Tyr Lys Met Thr Ile Ser Met Ala Asp Trp Pro Ser 450
455 460 Glu Ala Ser Glu Asp Trp
Ile Phe His Val Leu Ser Gln Glu Arg Asp 465 470
475 480 Gln Ser Thr Asn Ile Thr Leu Ser Arg Lys Gly
Ile Val Lys Leu Asn 485 490
495 Ile Tyr Phe Gln Glu Phe Asn Tyr Arg Thr Ile Glu Glu Ser Ala Ala
500 505 510 Asn Asn
Ile Val Trp Leu Leu Ser Asn Leu Gly Gly Gln Phe Gly Phe 515
520 525 Trp Met Gly Gly Ser Val Leu
Cys Leu Ile Glu Phe Gly Glu Ile Ile 530 535
540 Ile Asp Phe Val Trp Ile Thr Ile Ile Lys Leu Val
Ala Leu Ala Lys 545 550 555
560 Ser Leu Arg Gln Arg Arg Ala Gln Ala Ser Tyr Ala Gly Pro Pro Pro
565 570 575 Thr Val Ala
Glu Leu Val Glu Ala His Thr Asn Phe Gly Phe Gln Pro 580
585 590 Asp Thr Ala Pro Arg Ser Pro Asn
Thr Gly Pro Tyr Pro Ser Glu Gln 595 600
605 Ala Leu Pro Ile Pro Gly Thr Pro Pro Pro Asn Tyr Asp
Ser Leu Arg 610 615 620
Leu Gln Pro Leu Asp Val Ile Glu Ser Asp Ser Glu Gly Asp Ala Ile 625
630 635 640 51950DNAHomo
Sapiens 5atggcacccg gagagaagat caaagccaaa atcaagaaga atctgcccgt
gacgggccct 60caggcgccga ccattaaaga gctgatgcgg tggtactgcc tcaacaccaa
cacccatggc 120tgtcgccgca tcgtggtgtc ccgcggccgt ctgcgccgcc tcctctggat
cgggttcaca 180ctgactgccg tggccctcat cctctggcag tgcgccctcc tcgtcttctc
cttctatact 240gtctcagttt ccatcaaagt ccacttccgg aagctggatt ttcctgcagt
caccatctgc 300aacatcaacc cctacaagta cagcaccgtt cgccaccttc tagctgactt
ggaacaggag 360accagagagg ccctgaagtc cctgtatggc tttccagagt cccggaagcg
ccgagaggcg 420gagtcctgga actccgtctc agagggaaag cagcctagat tctcccaccg
gattccgctg 480ctgatctttg atcaggatga gaagggcaag gccagggact tcttcacagg
gaggaagcgg 540aaagtcggcg gtagcatcat tcacaaggct tcaaatgtca tgcacatcga
gtccaagcaa 600gtggtgggat tccaactgtg ctcaaatgac acctccgact gtgccaccta
caccttcagc 660tcgggaatca atgccattca ggagtggtat aagctacact acatgaacat
catggcacag 720gtgcctctgg agaagaaaat caacatgagc tattctgctg aggagctgct
ggtgacctgc 780ttctttgatg gagtgtcctg tgatgccagg aatttcacgc ttttccacca
cccgatgcat 840gggaattgct atactttcaa caacagagaa aatgagacca ttctcagcac
ctccatgggg 900ggcagcgaat atgggctgca agtcattttg tacataaacg aagaggaata
caacccattc 960ctcgtgtcct ccactggagc taaggtgatc atccatcggc aggatgagta
tcccttcgtc 1020gaagatgtgg gaacagagat tgagacagca atggtcacct ctataggaat
gcacctgaca 1080gagtccttca agctgagtga gccctacagt cagtgcacgg aggacgggag
tgacgtgcca 1140atcaggaaca tctacaacgc tgcctactcg ctccagatct gccttcattc
atgcttccag 1200acaaagatgg tggagaaatg tgggtgtgcc cagtacagcc agcctctacc
tcctgcagcc 1260aactactgca actaccagca gcaccccaac tggatgtatt gttactacca
actgcatcga 1320gcctttgtcc aggaagagct gggctgccag tctgtgtgca aggaagcctg
cagctttaaa 1380gagtggacac taaccacaag cctggcacaa tggccatctg tggtttcgga
gaagtggttg 1440ctgcctgttc tcacttggga ccaaggccgg caagtaaaca aaaagctcaa
caagacagac 1500ttggccaaac tcttgatatt ctacaaagac ctgaaccaga gatccatcat
ggagagccca 1560gccaacagta ttgagatgct tctgtccaac ttcggtggcc agctgggcct
gtggatgagc 1620tgctctgttg tctgcgtcat cgagatcatc gaggtcttct tcattgactt
cttctctatc 1680attgcccgcc gccagtggca gaaagccaag gagtggtggg cctggaaaca
ggctccccca 1740tgtccagaag ctccccgtag cccacagggc caggacaatc cagccctgga
tatagacgat 1800gacctaccca ctttcaactc tgctttgcac ctgcctccag ccctaggaac
ccaagtgccc 1860ggcacaccgc cccccaaata caataccttg cgcttggaga gggccttttc
caaccagctc 1920acagataccc agatgctaga tgagctctga
19506649PRTHomo Sapiens 6Met Ala Pro Gly Glu Lys Ile Lys Ala
Lys Ile Lys Lys Asn Leu Pro 1 5 10
15 Val Thr Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg
Trp Tyr 20 25 30
Cys Leu Asn Thr Asn Thr His Gly Cys Arg Arg Ile Val Val Ser Arg
35 40 45 Gly Arg Leu Arg
Arg Leu Leu Trp Ile Gly Phe Thr Leu Thr Ala Val 50
55 60 Ala Leu Ile Leu Trp Gln Cys Ala
Leu Leu Val Phe Ser Phe Tyr Thr 65 70
75 80 Val Ser Val Ser Ile Lys Val His Phe Arg Lys Leu
Asp Phe Pro Ala 85 90
95 Val Thr Ile Cys Asn Ile Asn Pro Tyr Lys Tyr Ser Thr Val Arg His
100 105 110 Leu Leu Ala
Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu Lys Ser Leu 115
120 125 Tyr Gly Phe Pro Glu Ser Arg Lys
Arg Arg Glu Ala Glu Ser Trp Asn 130 135
140 Ser Val Ser Glu Gly Lys Gln Pro Arg Phe Ser His Arg
Ile Pro Leu 145 150 155
160 Leu Ile Phe Asp Gln Asp Glu Lys Gly Lys Ala Arg Asp Phe Phe Thr
165 170 175 Gly Arg Lys Arg
Lys Val Gly Gly Ser Ile Ile His Lys Ala Ser Asn 180
185 190 Val Met His Ile Glu Ser Lys Gln Val
Val Gly Phe Gln Leu Cys Ser 195 200
205 Asn Asp Thr Ser Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly
Ile Asn 210 215 220
Ala Ile Gln Glu Trp Tyr Lys Leu His Tyr Met Asn Ile Met Ala Gln 225
230 235 240 Val Pro Leu Glu Lys
Lys Ile Asn Met Ser Tyr Ser Ala Glu Glu Leu 245
250 255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser
Cys Asp Ala Arg Asn Phe 260 265
270 Thr Leu Phe His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn
Asn 275 280 285 Arg
Glu Asn Glu Thr Ile Leu Ser Thr Ser Met Gly Gly Ser Glu Tyr 290
295 300 Gly Leu Gln Val Ile Leu
Tyr Ile Asn Glu Glu Glu Tyr Asn Pro Phe 305 310
315 320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile
His Arg Gln Asp Glu 325 330
335 Tyr Pro Phe Val Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val
340 345 350 Thr Ser
Ile Gly Met His Leu Thr Glu Ser Phe Lys Leu Ser Glu Pro 355
360 365 Tyr Ser Gln Cys Thr Glu Asp
Gly Ser Asp Val Pro Ile Arg Asn Ile 370 375
380 Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His
Ser Cys Phe Gln 385 390 395
400 Thr Lys Met Val Glu Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu
405 410 415 Pro Pro Ala
Ala Asn Tyr Cys Asn Tyr Gln Gln His Pro Asn Trp Met 420
425 430 Tyr Cys Tyr Tyr Gln Leu His Arg
Ala Phe Val Gln Glu Glu Leu Gly 435 440
445 Cys Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu
Trp Thr Leu 450 455 460
Thr Thr Ser Leu Ala Gln Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465
470 475 480 Leu Pro Val Leu
Thr Trp Asp Gln Gly Arg Gln Val Asn Lys Lys Leu 485
490 495 Asn Lys Thr Asp Leu Ala Lys Leu Leu
Ile Phe Tyr Lys Asp Leu Asn 500 505
510 Gln Arg Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met
Leu Leu 515 520 525
Ser Asn Phe Gly Gly Gln Leu Gly Leu Trp Met Ser Cys Ser Val Val 530
535 540 Cys Val Ile Glu Ile
Ile Glu Val Phe Phe Ile Asp Phe Phe Ser Ile 545 550
555 560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys
Glu Trp Trp Ala Trp Lys 565 570
575 Gln Ala Pro Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln
Asp 580 585 590 Asn
Pro Ala Leu Asp Ile Asp Asp Asp Leu Pro Thr Phe Asn Ser Ala 595
600 605 Leu His Leu Pro Pro Ala
Leu Gly Thr Gln Val Pro Gly Thr Pro Pro 610 615
620 Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala
Phe Ser Asn Gln Leu 625 630 635
640 Thr Asp Thr Gln Met Leu Asp Glu Leu 645
71917DNAHomo Sapiens 7atggctgagc accgaagcat ggacgggaga atggaagcag
ccacacgggg gggctctcac 60ctccaggctg cagcccagac gccccccagg ccggggccac
catcagcacc accaccacca 120cccaaggagg ggcaccagga ggggctggtg gagctgcccg
cctcgttccg ggagctgctc 180accttcttct gcaccaatgc caccatccac ggcgccatcc
gcctggtctg ctcccgcggg 240aaccgcctca agacgacgtc ctgggggctg ctgtccctgg
gagccctggt cgcgctctgc 300tggcagctgg ggctcctctt tgagcgtcac tggcaccgcc
cggtcctcat ggccgtctct 360gtgcactcgg agcgcaagct gctcccgctg gtcaccctgt
gtgacgggaa cccacgtcgg 420ccgagtccgg tcctccgcca tctggagctg ctggacgagt
ttgccaggga gaacattgac 480tccctgtaca acgtcaacct cagcaaaggc agagccgccc
tctccgccac tgtcccccgc 540cacgagcccc ccttccacct ggaccgggag atccgtctgc
agaggctgag ccactcgggc 600agccgggtca gagtggggtt cagactgtgc aacagcacgg
gcggcgactg cttttaccga 660ggctacacgt caggcgtggc ggctgtccag gactggtacc
acttccacta tgtggatatc 720ctggccctgc tgcccgcggc atgggaggac agccacggga
gccaggacgg ccacttcgtc 780ctctcctgca gttacgatgg cctggactgc caggcccgac
agttccggac cttccaccac 840cccacctacg gcagctgcta cacggtcgat ggcgtctgga
cagctcagcg ccccggcatc 900acccacggag tcggcctggt cctcagggtt gagcagcagc
ctcacctccc tctgctgtcc 960acgctggccg gcatcagggt catggttcac ggccgtaacc
acacgccctt cctggggcac 1020cacagcttca gcgtccggcc agggacggag gccaccatca
gcatccgaga ggacgaggtg 1080caccggctcg ggagccccta cggccactgc accgccggcg
gggaaggcgt ggaggtggag 1140ctgctacaca acacctccta caccaggcag gcctgcctgg
tgtcctgctt ccagcagctg 1200atggtggaga cctgctcctg tggctactac ctccaccctc
tgccggcggg ggctgagtac 1260tgcagctctg cccggcaccc tgcctgggga cactgcttct
accgcctcta ccaggacctg 1320gagacccacc ggctcccctg tacctcccgc tgccccaggc
cctgcaggga gtctgcattc 1380aagctctcca ctgggacctc caggtggcct tccgccaagt
cagctggatg gactctggcc 1440acgctaggtg aacaggggct gccgcatcag agccacagac
agaggagcag cctggccaaa 1500atcaacatcg tctaccagga gctcaactac cgctcagtgg
aggaggcgcc cgtgtactcg 1560gtgccgcagc tgctctccgc catgggcagc ctctacagcc
tgtggtttgg ggcctccgtc 1620ctctccctcc tggagctcct ggagctgctg ctcgatgctt
ctgccctcac cctggtgcta 1680ggcggccgcc ggctccgcag ggcgtggttc tcctggccca
gagccagccc tgcctcaggg 1740gcgtccagca tcaagccaga ggccagtcag atgcccccgc
ctgcaggcgg cacgtcagat 1800gacccggagc ccagcgggcc tcatctccca cgggtgatgc
ttccaggggt tctggcggga 1860gtctcagccg aagagagctg ggctgggccc cagccccttg
agactctgga cacctga 19178638PRTHomo Sapiens 8Met Ala Glu His Arg Ser
Met Asp Gly Arg Met Glu Ala Ala Thr Arg 1 5
10 15 Gly Gly Ser His Leu Gln Ala Ala Ala Gln Thr
Pro Pro Arg Pro Gly 20 25
30 Pro Pro Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly His Gln Glu
Gly 35 40 45 Leu
Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu Thr Phe Phe Cys 50
55 60 Thr Asn Ala Thr Ile His
Gly Ala Ile Arg Leu Val Cys Ser Arg Gly 65 70
75 80 Asn Arg Leu Lys Thr Thr Ser Trp Gly Leu Leu
Ser Leu Gly Ala Leu 85 90
95 Val Ala Leu Cys Trp Gln Leu Gly Leu Leu Phe Glu Arg His Trp His
100 105 110 Arg Pro
Val Leu Met Ala Val Ser Val His Ser Glu Arg Lys Leu Leu 115
120 125 Pro Leu Val Thr Leu Cys Asp
Gly Asn Pro Arg Arg Pro Ser Pro Val 130 135
140 Leu Arg His Leu Glu Leu Leu Asp Glu Phe Ala Arg
Glu Asn Ile Asp 145 150 155
160 Ser Leu Tyr Asn Val Asn Leu Ser Lys Gly Arg Ala Ala Leu Ser Ala
165 170 175 Thr Val Pro
Arg His Glu Pro Pro Phe His Leu Asp Arg Glu Ile Arg 180
185 190 Leu Gln Arg Leu Ser His Ser Gly
Ser Arg Val Arg Val Gly Phe Arg 195 200
205 Leu Cys Asn Ser Thr Gly Gly Asp Cys Phe Tyr Arg Gly
Tyr Thr Ser 210 215 220
Gly Val Ala Ala Val Gln Asp Trp Tyr His Phe His Tyr Val Asp Ile 225
230 235 240 Leu Ala Leu Leu
Pro Ala Ala Trp Glu Asp Ser His Gly Ser Gln Asp 245
250 255 Gly His Phe Val Leu Ser Cys Ser Tyr
Asp Gly Leu Asp Cys Gln Ala 260 265
270 Arg Gln Phe Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys
Tyr Thr 275 280 285
Val Asp Gly Val Trp Thr Ala Gln Arg Pro Gly Ile Thr His Gly Val 290
295 300 Gly Leu Val Leu Arg
Val Glu Gln Gln Pro His Leu Pro Leu Leu Ser 305 310
315 320 Thr Leu Ala Gly Ile Arg Val Met Val His
Gly Arg Asn His Thr Pro 325 330
335 Phe Leu Gly His His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala
Thr 340 345 350 Ile
Ser Ile Arg Glu Asp Glu Val His Arg Leu Gly Ser Pro Tyr Gly 355
360 365 His Cys Thr Ala Gly Gly
Glu Gly Val Glu Val Glu Leu Leu His Asn 370 375
380 Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser
Cys Phe Gln Gln Leu 385 390 395
400 Met Val Glu Thr Cys Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala
405 410 415 Gly Ala
Glu Tyr Cys Ser Ser Ala Arg His Pro Ala Trp Gly His Cys 420
425 430 Phe Tyr Arg Leu Tyr Gln Asp
Leu Glu Thr His Arg Leu Pro Cys Thr 435 440
445 Ser Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe
Lys Leu Ser Thr 450 455 460
Gly Thr Ser Arg Trp Pro Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465
470 475 480 Thr Leu Gly
Glu Gln Gly Leu Pro His Gln Ser His Arg Gln Arg Ser 485
490 495 Ser Leu Ala Lys Ile Asn Ile Val
Tyr Gln Glu Leu Asn Tyr Arg Ser 500 505
510 Val Glu Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu
Ser Ala Met 515 520 525
Gly Ser Leu Tyr Ser Leu Trp Phe Gly Ala Ser Val Leu Ser Leu Leu 530
535 540 Glu Leu Leu Glu
Leu Leu Leu Asp Ala Ser Ala Leu Thr Leu Val Leu 545 550
555 560 Gly Gly Arg Arg Leu Arg Arg Ala Trp
Phe Ser Trp Pro Arg Ala Ser 565 570
575 Pro Ala Ser Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln
Met Pro 580 585 590
Pro Pro Ala Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser Gly Pro His
595 600 605 Leu Pro Arg Val
Met Leu Pro Gly Val Leu Ala Gly Val Ser Ala Glu 610
615 620 Glu Ser Trp Ala Gly Pro Gln Pro
Leu Glu Thr Leu Asp Thr 625 630 635
9638PRTHomo Sapiens 9Met Ala Glu His Arg Ser Met Asp Gly Arg Met
Glu Ala Ala Thr Arg 1 5 10
15 Gly Gly Ser His Leu Gln Ala Ala Ala Gln Thr Pro Pro Arg Pro Gly
20 25 30 Pro Pro
Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly His Gln Glu Gly 35
40 45 Leu Val Glu Leu Pro Ala Ser
Phe Arg Glu Leu Leu Thr Phe Phe Cys 50 55
60 Thr Asn Ala Thr Ile His Gly Ala Ile Arg Leu Val
Cys Ser Arg Gly 65 70 75
80 Asn Arg Leu Lys Thr Thr Ser Trp Gly Leu Leu Ser Leu Gly Ala Leu
85 90 95 Val Ala Leu
Cys Trp Gln Leu Gly Leu Leu Phe Glu Arg His Trp His 100
105 110 Arg Pro Val Leu Met Ala Val Ser
Val His Ser Glu Arg Lys Leu Leu 115 120
125 Pro Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro
Ser Pro Val 130 135 140
Leu Arg His Leu Glu Leu Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145
150 155 160 Ser Leu Tyr Asn
Val Asn Leu Ser Lys Gly Arg Ala Ala Leu Ser Ala 165
170 175 Thr Val Pro Arg His Glu Pro Pro Phe
His Leu Asp Arg Glu Ile Arg 180 185
190 Leu Gln Arg Leu Ser His Ser Gly Ser Arg Val Arg Val Gly
Phe Arg 195 200 205
Leu Cys Asn Ser Thr Gly Gly Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210
215 220 Gly Val Ala Ala Val
Gln Asp Trp Tyr His Phe His Tyr Val Asp Ile 225 230
235 240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp
Ser His Gly Ser Gln Asp 245 250
255 Gly His Phe Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln
Ala 260 265 270 Arg
Gln Phe Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys Tyr Thr 275
280 285 Val Asp Gly Val Trp Thr
Ala Gln Arg Pro Gly Ile Thr His Gly Val 290 295
300 Gly Leu Val Leu Arg Val Glu Gln Gln Pro His
Leu Pro Leu Leu Ser 305 310 315
320 Thr Leu Ala Gly Ile Arg Val Met Val His Gly Arg Asn His Thr Pro
325 330 335 Phe Leu
Gly His His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala Thr 340
345 350 Ile Ser Ile Arg Glu Asp Glu
Val His Arg Leu Gly Ser Pro Tyr Gly 355 360
365 His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Glu
Leu Leu His Asn 370 375 380
Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser Cys Phe Gln Gln Leu 385
390 395 400 Met Val Glu
Thr Cys Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala 405
410 415 Gly Ala Glu Tyr Cys Ser Ser Ala
Arg His Pro Ala Trp Gly His Cys 420 425
430 Phe Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu
Pro Cys Thr 435 440 445
Ser Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450
455 460 Gly Thr Ser Arg
Trp Pro Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465 470
475 480 Thr Leu Gly Glu Gln Gly Leu Pro His
Gln Ser His Arg Gln Arg Ser 485 490
495 Ser Leu Ala Lys Ile Asn Ile Val Tyr Gln Glu Leu Asn Tyr
Arg Ser 500 505 510
Val Glu Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu Ser Ala Met
515 520 525 Gly Ser Leu Cys
Ser Leu Trp Phe Gly Ala Ser Val Leu Ser Leu Leu 530
535 540 Glu Leu Leu Glu Leu Leu Leu Asp
Ala Ser Ala Leu Thr Leu Val Leu 545 550
555 560 Gly Gly Arg Arg Leu Arg Arg Ala Trp Phe Ser Trp
Pro Arg Ala Ser 565 570
575 Pro Ala Ser Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln Met Pro
580 585 590 Pro Pro Ala
Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser Gly Pro His 595
600 605 Leu Pro Arg Val Met Leu Pro Gly
Val Leu Ala Gly Val Ser Ala Glu 610 615
620 Glu Ser Trp Ala Gly Pro Gln Pro Leu Glu Thr Leu Asp
Thr 625 630 635 1011PRTHomo
Sapiens 10Met Gly Ser Gln Trp Ser Leu Trp Phe Gly Ala 1 5
10 1111PRTHomo Sapiens 11Met Gly Ser Leu Tyr Ser Leu
Trp Phe Gly Ala 1 5 10 1211PRTHomo
Sapiens 12Met Gly Ser Leu Cys Ser Leu Trp Phe Gly Ala 1 5
10 13649PRTHomo Sapiens 13Met Ala Pro Gly Glu Lys
Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro 1 5
10 15 Val Thr Gly Pro Gln Ala Pro Thr Ile Lys Glu
Leu Met Arg Trp Tyr 20 25
30 Cys Leu Asn Thr Asn Thr His Gly Cys Arg Arg Ile Val Val Ser
Arg 35 40 45 Gly
Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr Leu Thr Ala Val 50
55 60 Ala Leu Ile Leu Trp Gln
Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65 70
75 80 Val Ser Val Ser Ile Lys Val His Phe Arg Lys
Leu Asp Phe Pro Ala 85 90
95 Val Thr Ile Cys Asn Ile Asn Pro Tyr Lys Tyr Ser Thr Val Arg His
100 105 110 Leu Leu
Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu Lys Ser Leu 115
120 125 Tyr Gly Phe Pro Glu Ser Arg
Lys Arg Arg Glu Ala Glu Ser Trp Asn 130 135
140 Ser Val Ser Glu Gly Lys Gln Pro Arg Phe Ser His
Arg Ile Pro Pro 145 150 155
160 Leu Ile Phe Asp Gln Asp Glu Lys Gly Lys Ala Arg Asp Phe Phe Thr
165 170 175 Gly Arg Lys
Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala Ser Asn 180
185 190 Val Met His Ile Glu Ser Lys Gln
Val Val Gly Phe Gln Leu Cys Ser 195 200
205 Asn Asp Thr Ser Asp Cys Ala Thr Tyr Thr Phe Ser Leu
Gly Ile Asn 210 215 220
Ala Ile Gln Glu Trp Tyr Lys Leu His Tyr Met Asn Ile Met Ala Gln 225
230 235 240 Val Pro Leu Glu
Lys Lys Ile Asn Met Ser Tyr Ser Ala Glu Glu Leu 245
250 255 Leu Val Thr Cys Phe Phe Asp Gly Val
Ser Cys Asp Ala Arg Asn Phe 260 265
270 Thr Leu Phe His His Pro Met His Gly Asn Cys Tyr Thr Phe
Asn Asn 275 280 285
Arg Glu Asn Glu Thr Ile Leu Ser Thr Ser Met Gly Gly Ser Glu Tyr 290
295 300 Gly Leu Gln Val Ile
Leu Tyr Ile Asn Glu Glu Glu Tyr Asn Pro Phe 305 310
315 320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile
Ile His Arg Gln Asp Glu 325 330
335 Tyr Pro Phe Val Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met
Val 340 345 350 Thr
Ser Ile Gly Met His Leu Thr Glu Ser Phe Lys Leu Ser Glu Pro 355
360 365 Tyr Ser Gln Cys Thr Glu
Asp Gly Ser Asp Val Pro Ile Arg Asn Ile 370 375
380 Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu
His Ser Cys Phe Gln 385 390 395
400 Thr Lys Met Val Glu Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu
405 410 415 Pro Pro
Ala Ala Asn Tyr Cys Asn Tyr Gln Gln His Pro Asn Trp Met 420
425 430 Tyr Cys Tyr Tyr Gln Leu His
Arg Ala Phe Val Gln Glu Glu Leu Gly 435 440
445 Cys Gln Ser Val Cys Lys Glu Ala Cys Ser Leu Lys
Glu Trp Thr Leu 450 455 460
Thr Thr Ser Leu Ala Gln Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465
470 475 480 Leu Pro Val
Leu Thr Trp Asp Gln Gly Arg Gln Val Asn Lys Lys Leu 485
490 495 Asn Lys Thr Asp Leu Ala Lys Leu
Leu Ile Phe Tyr Lys Asp Leu Asn 500 505
510 Gln Arg Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu
Met Leu Leu 515 520 525
Ser Asn Phe Gly Gly Gln Leu Gly Leu Trp Met Ser Cys Ser Val Val 530
535 540 Cys Val Val Glu
Ile Ile Glu Val Phe Phe Ile Asp Phe Phe Ser Ile 545 550
555 560 Ile Ala Arg Arg Gln Trp Gln Lys Ala
Lys Glu Trp Trp Ala Trp Lys 565 570
575 Gln Ala Pro Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly
Gln Asp 580 585 590
Asn Pro Ala Leu Asp Ile Asp Asp Asp Leu Pro Thr Phe Asn Ser Ala
595 600 605 Leu His Leu Pro
Pro Ala Leu Gly Thr Gln Val Pro Gly Thr Pro Pro 610
615 620 Pro Lys Tyr Asn Thr Leu Arg Leu
Glu Arg Ala Phe Ser Asn Gln Leu 625 630
635 640 Thr Asp Thr Gln Met Leu Asp Glu Leu
645 14649PRTHomo Sapiens 14Met Ala Pro Gly Glu Lys Ile
Lys Ala Lys Ile Lys Lys Asn Leu Pro 1 5
10 15 Val Thr Gly Pro Gln Ala Pro Thr Ile Lys Glu
Leu Met Arg Trp Tyr 20 25
30 Cys Leu Asn Thr Asn Thr His Gly Cys Arg Arg Ile Val Val Ser
Arg 35 40 45 Gly
Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr Leu Thr Ala Val 50
55 60 Ala Leu Ile Leu Trp Gln
Cys Ala Leu Leu Val Phe Ser Phe Tyr Ala 65 70
75 80 Val Ser Val Ser Ile Lys Val His Phe Arg Lys
Leu Asp Phe Pro Ala 85 90
95 Val Thr Ile Cys Asn Ile Asn Pro Tyr Lys Tyr Ser Thr Val Arg His
100 105 110 Leu Leu
Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu Lys Ser Leu 115
120 125 Tyr Gly Phe Pro Glu Ser Arg
Lys Arg Arg Glu Ala Glu Ser Trp Asn 130 135
140 Ser Val Ser Glu Gly Lys Gln Pro Arg Phe Ser His
Arg Ile Pro Leu 145 150 155
160 Leu Ile Phe Asp Gln Asp Glu Lys Gly Lys Ala Arg Asp Phe Phe Thr
165 170 175 Gly Arg Lys
Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala Ser Asn 180
185 190 Val Met His Ile Glu Ser Lys Gln
Val Val Gly Phe Gln Leu Cys Ser 195 200
205 Asn Asp Thr Ser Asp Cys Ala Thr Tyr Thr Phe Ser Ser
Gly Ile Asn 210 215 220
Ala Ile Gln Glu Trp Tyr Lys Leu His Tyr Met Asn Ile Met Ala Gln 225
230 235 240 Val Pro Leu Glu
Lys Lys Ile Asn Met Ser Tyr Ser Ala Glu Glu Leu 245
250 255 Leu Val Thr Cys Phe Phe Asp Gly Val
Ser Cys Asp Ala Arg Asn Phe 260 265
270 Thr Leu Phe His His Pro Met His Gly Asn Cys Tyr Thr Phe
Asn Asn 275 280 285
Arg Glu Asn Glu Thr Ile Leu Ser Thr Ser Met Gly Gly Ser Glu Tyr 290
295 300 Gly Leu Gln Val Ile
Leu Tyr Ile Asn Glu Glu Glu Tyr Asn Pro Phe 305 310
315 320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile
Ile His Arg Gln Asp Glu 325 330
335 Tyr Pro Phe Val Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met
Val 340 345 350 Thr
Ser Ile Gly Met His Leu Thr Glu Ser Phe Lys Leu Ser Glu Pro 355
360 365 Tyr Ser Gln Cys Thr Glu
Asp Gly Ser Asp Val Pro Ile Arg Asn Ile 370 375
380 Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu
His Ser Cys Phe Gln 385 390 395
400 Thr Lys Met Val Glu Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu
405 410 415 Pro Pro
Ala Ala Asn Tyr Cys Asn Tyr Gln Gln His Pro Asn Trp Met 420
425 430 Tyr Cys Tyr Tyr Gln Leu His
Arg Ala Phe Val Gln Glu Glu Leu Gly 435 440
445 Cys Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys
Glu Trp Thr Leu 450 455 460
Thr Thr Ser Leu Ala Gln Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465
470 475 480 Leu Pro Val
Leu Thr Trp Asp Gln Gly Arg Gln Val Asn Lys Lys Leu 485
490 495 Asn Lys Thr Asp Leu Ala Lys Leu
Leu Ile Phe Tyr Lys Asp Leu Asn 500 505
510 Gln Arg Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu
Met Leu Leu 515 520 525
Ser Asn Phe Gly Gly Gln Leu Gly Leu Trp Met Ser Cys Ser Val Val 530
535 540 Cys Val Ile Glu
Ile Ile Glu Val Phe Phe Ile Asp Phe Phe Ser Ile 545 550
555 560 Ile Ala Arg Arg Gln Trp Gln Lys Ala
Lys Glu Trp Trp Ala Trp Lys 565 570
575 Gln Ala Pro Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly
Gln Asp 580 585 590
Asn Pro Ala Leu Asp Ile Asp Asp Asp Leu Pro Thr Phe Asn Ser Ala
595 600 605 Leu His Leu Pro
Pro Ala Leu Gly Thr Gln Val Pro Gly Thr Pro Pro 610
615 620 Pro Lys Tyr Asn Thr Leu Arg Leu
Glu Arg Ala Phe Ser Asn Ser Leu 625 630
635 640 Thr Asp Thr Gln Met Leu Asp Glu Leu
645 15649PRTHomo Sapiens 15Met Ala Pro Gly Glu Lys Ile
Lys Ala Lys Ile Lys Lys Asn Leu Pro 1 5
10 15 Val Thr Gly Pro Gln Ala Pro Thr Ile Lys Glu
Leu Met Arg Trp Tyr 20 25
30 Cys Leu Asn Thr Asn Thr His Gly Cys Arg Arg Ile Val Val Ser
Arg 35 40 45 Gly
Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr Leu Thr Ala Val 50
55 60 Ala Leu Ile Leu Trp Gln
Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65 70
75 80 Val Ser Val Ser Ile Lys Val His Phe Arg Lys
Leu Asp Phe Pro Ala 85 90
95 Val Thr Ile Cys Asn Ile Asn Pro Tyr Lys Tyr Ser Thr Val Arg His
100 105 110 Leu Leu
Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu Lys Ser Leu 115
120 125 Tyr Gly Phe Pro Glu Ser Arg
Lys Arg Arg Glu Ala Glu Ser Trp Asn 130 135
140 Ser Val Ser Glu Gly Lys Gln Pro Arg Phe Ser His
Arg Ile Pro Leu 145 150 155
160 Leu Ile Phe Asp Gln Asp Glu Lys Gly Lys Ala Arg Asp Phe Phe Thr
165 170 175 Gly Arg Lys
Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala Ser Asn 180
185 190 Val Met His Ile Glu Ser Lys Gln
Val Val Gly Phe Gln Leu Cys Ser 195 200
205 Asn Asp Thr Ser Asp Cys Ala Thr Tyr Thr Phe Ser Ser
Gly Ile Asn 210 215 220
Ala Ile Gln Glu Trp Tyr Lys Leu His Tyr Met Asn Ile Met Ala Gln 225
230 235 240 Val Pro Leu Glu
Lys Lys Ile Asn Met Ser Tyr Ser Ala Glu Glu Leu 245
250 255 Leu Val Thr Cys Phe Phe Asp Gly Val
Ser Cys Asp Ala Arg Asn Phe 260 265
270 Thr Leu Phe His His Pro Met His Gly Asn Cys Tyr Thr Phe
Asn Asn 275 280 285
Arg Glu Asn Glu Thr Ile Leu Ser Thr Ser Met Gly Gly Ser Glu Tyr 290
295 300 Gly Leu Gln Val Ile
Leu Tyr Ile Asn Glu Glu Glu Tyr Asn Pro Phe 305 310
315 320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile
Ile His Arg Gln Asp Glu 325 330
335 Tyr Pro Phe Val Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met
Val 340 345 350 Thr
Ser Ile Gly Met His Leu Thr Glu Ser Phe Lys Leu Ser Glu Pro 355
360 365 Tyr Ser Gln Cys Thr Glu
Asp Gly Ser Asp Val Pro Ile Arg Asn Ile 370 375
380 Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu
His Ser Cys Phe Gln 385 390 395
400 Thr Lys Met Val Glu Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu
405 410 415 Pro Pro
Ala Ala Asn Tyr Cys Asn Tyr Gln Gln His Pro Asn Trp Met 420
425 430 Tyr Cys Tyr Tyr Gln Leu His
Arg Ala Phe Val Gln Glu Glu Leu Gly 435 440
445 Cys Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys
Glu Trp Thr Leu 450 455 460
Thr Thr Ser Leu Ala Gln Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465
470 475 480 Leu Pro Val
Leu Thr Trp Asp Gln Gly Arg Gln Val Asn Lys Lys Leu 485
490 495 Asn Lys Thr Asp Leu Ala Lys Leu
Leu Ile Phe Tyr Lys Asp Leu Asn 500 505
510 Gln Arg Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu
Met Leu Leu 515 520 525
Ser Asn Phe Gly Gly Gln Leu Gly Leu Trp Met Ser Cys Ser Val Val 530
535 540 Cys Val Ile Glu
Ile Ile Glu Val Phe Phe Ile Asp Phe Phe Ser Ile 545 550
555 560 Ile Ala Arg Arg Gln Trp Gln Lys Ala
Lys Glu Trp Trp Ala Trp Lys 565 570
575 Gln Ala Pro Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly
Gln Asp 580 585 590
Asn Pro Ala Leu Asp Ile Asp Asp Gly Leu Pro Thr Phe Asn Ser Ala
595 600 605 Leu His Leu Pro
Pro Ala Leu Gly Thr Gln Val Pro Gly Thr Pro Pro 610
615 620 Pro Lys Tyr Asn Thr Leu Arg Leu
Glu Arg Ala Phe Ser Asn Gln Leu 625 630
635 640 Thr Asp Thr Gln Met Leu Asp Glu Leu
645 16649PRTHomo Sapiens 16Met Ala Pro Gly Glu Lys Ile
Lys Ala Lys Ile Lys Lys Asn Leu Pro 1 5
10 15 Val Thr Gly Pro Gln Ala Pro Thr Ile Lys Glu
Leu Met Arg Trp Tyr 20 25
30 Cys Leu Asn Thr Asn Thr His Gly Cys Arg Arg Ile Val Val Ser
Arg 35 40 45 Gly
Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr Leu Thr Ala Val 50
55 60 Ala Leu Ile Leu Trp Gln
Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65 70
75 80 Val Ser Val Ser Ile Lys Val His Phe Arg Lys
Leu Asp Phe Pro Ala 85 90
95 Val Thr Ile Cys Asn Ile Asn Pro Tyr Lys Tyr Ser Thr Val Arg His
100 105 110 Leu Leu
Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu Lys Ser Leu 115
120 125 Tyr Gly Phe Pro Glu Ser Arg
Lys Arg Arg Glu Ala Glu Ser Trp Asn 130 135
140 Ser Val Ser Glu Gly Lys Gln Pro Arg Phe Ser His
Arg Ile Pro Leu 145 150 155
160 Leu Ile Phe Asp Gln Asp Glu Lys Gly Lys Ala Arg Asp Phe Phe Thr
165 170 175 Gly Arg Lys
Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala Ser Asn 180
185 190 Val Met His Ile Glu Ser Lys Gln
Val Val Gly Phe Gln Leu Cys Ser 195 200
205 Asn Asp Thr Ser Asp Cys Ala Thr Tyr Thr Phe Ser Ser
Gly Ile Asn 210 215 220
Ala Ile Gln Glu Trp Tyr Lys Leu His Tyr Met Asn Ile Met Ala Gln 225
230 235 240 Val Pro Leu Glu
Lys Lys Ile Asn Met Ser Tyr Ser Ala Glu Glu Leu 245
250 255 Leu Val Thr Cys Phe Phe Asp Gly Val
Ser Cys Asp Ala Arg Asn Phe 260 265
270 Thr Leu Phe His His Pro Met His Gly Asn Cys Tyr Thr Phe
Asn Asn 275 280 285
Arg Glu Asn Glu Thr Ile Leu Ser Thr Ser Met Gly Gly Ser Glu Tyr 290
295 300 Gly Leu Gln Val Ile
Leu Tyr Ile Asn Glu Glu Glu Tyr Asn Pro Phe 305 310
315 320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile
Ile His Arg Gln Asp Glu 325 330
335 Tyr Pro Ser Val Glu Asp Val Gly Thr Glu Ile Glu Thr Thr Met
Val 340 345 350 Thr
Ser Ile Gly Met His Leu Thr Glu Ser Phe Lys Leu Ser Glu Pro 355
360 365 Ser Ser Gln Cys Thr Glu
Gly Gly Ser Asp Val Pro Ile Arg Asn Ile 370 375
380 Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu
His Ser Cys Phe Gln 385 390 395
400 Thr Lys Met Val Glu Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu
405 410 415 Pro Pro
Ala Ala Asn Tyr Cys Asn Tyr Gln Gln His Pro Asn Trp Met 420
425 430 Tyr Cys Tyr Tyr Gln Leu His
Arg Ala Phe Val Gln Glu Glu Leu Gly 435 440
445 Cys Gln Ser Val Cys Lys Glu Ala Cys Arg Phe Lys
Glu Trp Thr Leu 450 455 460
Thr Thr Ser Leu Ala Gln Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465
470 475 480 Leu Pro Val
Leu Thr Trp Asp Gln Gly Arg Gln Val Asn Lys Lys Leu 485
490 495 Asn Lys Thr Asp Leu Ala Lys Leu
Leu Ile Phe Tyr Lys Asp Leu Asn 500 505
510 Gln Arg Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu
Met Leu Leu 515 520 525
Ser Asn Phe Gly Gly Gln Leu Gly Leu Trp Met Ser Cys Ser Val Val 530
535 540 Cys Val Ile Glu
Ile Ile Glu Val Phe Phe Ile Asp Phe Phe Ser Ile 545 550
555 560 Ile Ala Arg Arg Gln Trp Gln Lys Ala
Lys Glu Trp Trp Ala Trp Lys 565 570
575 Gln Ala Pro Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly
Gln Asp 580 585 590
Asn Pro Ala Leu Asp Ile Asp Asp Asp Leu Pro Thr Phe Asn Ser Ala
595 600 605 Leu His Leu Pro
Pro Ala Leu Gly Thr Gln Val Pro Gly Thr Pro Pro 610
615 620 Pro Lys Tyr Asn Thr Leu Arg Leu
Glu Arg Ala Phe Ser Asn Gln Leu 625 630
635 640 Thr Asp Thr Gln Met Leu Asp Glu Leu
645 17638PRTHomo Sapiens 17Met Ala Glu His Arg Ser Met
Asp Gly Arg Met Glu Ala Ala Thr Arg 1 5
10 15 Gly Gly Ser His Leu Gln Ala Ala Ala Gln Thr
Pro Pro Arg Pro Gly 20 25
30 Pro Pro Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly His Gln Glu
Gly 35 40 45 Leu
Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu Thr Phe Phe Cys 50
55 60 Thr Asn Ala Thr Ile His
Gly Ala Ile Arg Leu Val Cys Ser Arg Gly 65 70
75 80 Asn Arg Leu Lys Thr Thr Ser Trp Gly Leu Leu
Ser Leu Gly Ala Leu 85 90
95 Val Ala Leu Cys Trp Gln Leu Gly Leu Leu Phe Glu Arg His Trp His
100 105 110 Arg Pro
Val Leu Met Ala Val Ser Val His Ser Glu Arg Lys Leu Leu 115
120 125 Pro Leu Val Thr Leu Cys Asp
Gly Asn Pro Arg Arg Pro Ser Pro Val 130 135
140 Leu Arg His Leu Glu Leu Leu Asp Glu Phe Ala Arg
Glu Asn Ile Asp 145 150 155
160 Ser Leu Tyr Asn Val Asn Leu Ser Lys Gly Arg Ala Ala Leu Ser Ala
165 170 175 Thr Val Pro
Arg His Glu Pro Pro Phe His Leu Asp Arg Glu Ile Arg 180
185 190 Leu Gln Arg Leu Ser His Ser Gly
Ser Arg Val Arg Val Gly Phe Arg 195 200
205 Leu Cys Asn Ser Thr Gly Gly Asp Cys Phe Tyr Arg Gly
Tyr Thr Ser 210 215 220
Gly Val Ala Ala Val Gln Asp Trp Tyr His Phe His Tyr Val Asp Ile 225
230 235 240 Leu Ala Leu Leu
Pro Ala Ala Trp Glu Asp Ser His Gly Ser Gln Asp 245
250 255 Gly His Phe Val Leu Ser Cys Ser Tyr
Asp Gly Leu Asp Cys Gln Ala 260 265
270 Arg Gln Phe Arg Thr Ile His His Pro Thr Tyr Gly Ser Cys
Tyr Thr 275 280 285
Val Asp Gly Val Trp Thr Ala Gln Arg Pro Gly Ile Thr His Gly Val 290
295 300 Gly Leu Val Leu Arg
Val Glu Gln Gln Pro His Leu Pro Leu Leu Ser 305 310
315 320 Thr Leu Ala Gly Ile Arg Val Met Val His
Gly Arg Asn His Thr Pro 325 330
335 Phe Leu Gly His His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala
Thr 340 345 350 Ile
Arg Ile Arg Glu Asp Glu Val His Arg Leu Gly Ser Pro Tyr Gly 355
360 365 His Cys Thr Ala Gly Gly
Glu Gly Val Glu Val Glu Leu Leu His Asn 370 375
380 Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser
Cys Phe Gln Gln Leu 385 390 395
400 Met Val Glu Thr Cys Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala
405 410 415 Gly Ala
Glu Tyr Cys Ser Ser Ala Arg His Pro Ala Trp Gly His Cys 420
425 430 Phe Tyr Arg Leu Tyr Gln Asp
Leu Glu Thr His Arg Leu Pro Cys Thr 435 440
445 Ser Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe
Lys Leu Ser Thr 450 455 460
Gly Thr Ser Arg Trp Pro Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465
470 475 480 Thr Leu Gly
Glu Gln Gly Leu Pro His Gln Ser His Arg Gln Arg Ser 485
490 495 Ser Leu Ala Lys Ile Asn Ile Val
Tyr Gln Glu Leu Asn Tyr Arg Ser 500 505
510 Val Glu Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu
Ser Ala Met 515 520 525
Gly Ser Leu Cys Ser Leu Trp Phe Gly Ala Ser Val Leu Ser Leu Leu 530
535 540 Glu Leu Leu Glu
Leu Leu Leu Asp Ala Ser Ala Leu Thr Leu Val Leu 545 550
555 560 Gly Gly Arg Arg Leu His Arg Ala Trp
Phe Ser Trp Pro Arg Ala Ser 565 570
575 Pro Ala Ser Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln
Met Pro 580 585 590
Pro Pro Ala Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser Gly Pro His
595 600 605 Leu Pro Arg Val
Met Leu Pro Gly Val Leu Ala Gly Val Ser Ala Glu 610
615 620 Glu Ser Trp Ala Gly Pro Gln Pro
Leu Glu Thr Leu Asp Thr 625 630 635
18638PRTHomo Sapiens 18Met Ala Glu His Arg Ser Met Asp Gly Arg Met
Glu Ala Ala Thr Arg 1 5 10
15 Gly Gly Ser His Leu Gln Ala Ala Ala Gln Thr Pro Pro Arg Pro Gly
20 25 30 Pro Pro
Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly His Gln Glu Gly 35
40 45 Leu Val Glu Leu Pro Ala Ser
Phe Arg Glu Leu Leu Thr Phe Phe Cys 50 55
60 Thr Asn Ala Thr Ile His Gly Ala Ile Arg Leu Val
Cys Ser Arg Gly 65 70 75
80 Asn Arg Leu Lys Thr Thr Ser Trp Gly Leu Leu Ser Leu Gly Ala Leu
85 90 95 Val Ala Leu
Cys Trp Gln Leu Gly Leu Leu Phe Glu Arg His Trp His 100
105 110 Arg Pro Val Leu Met Ala Val Ser
Val His Ser Glu Arg Lys Leu Leu 115 120
125 Pro Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro
Ser Pro Val 130 135 140
Leu Arg His Leu Glu Leu Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145
150 155 160 Ser Leu Tyr Asn
Val Asn Leu Ser Lys Gly Arg Ala Ala Leu Ser Ala 165
170 175 Thr Val Pro Arg His Glu Pro Pro Phe
His Leu Asp Arg Glu Ile Arg 180 185
190 Leu Gln Arg Leu Ser His Ser Gly Ser Arg Val Arg Val Gly
Phe Arg 195 200 205
Leu Cys Asn Ser Thr Gly Gly Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210
215 220 Gly Val Ala Ala Val
Gln Asp Trp Tyr His Phe His Tyr Val Asp Ile 225 230
235 240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp
Ser His Gly Ser Gln Asp 245 250
255 Gly His Phe Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln
Ala 260 265 270 Arg
Gln Phe Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys Tyr Thr 275
280 285 Val Asp Gly Val Trp Thr
Ala Gln Arg Pro Gly Ile Thr His Gly Val 290 295
300 Gly Leu Val Leu Arg Val Glu Gln Gln Pro His
Leu Pro Leu Leu Ser 305 310 315
320 Thr Leu Ala Gly Ile Arg Val Met Val His Gly Arg Asn His Thr Pro
325 330 335 Phe Leu
Gly His His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala Thr 340
345 350 Ile Ser Ile Arg Glu Asp Glu
Val His Arg Leu Gly Ser Pro Tyr Gly 355 360
365 His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Glu
Leu Leu His Asn 370 375 380
Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser Cys Phe Gln Gln Leu 385
390 395 400 Met Val Glu
Thr Cys Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala 405
410 415 Gly Ala Glu Tyr Cys Ser Ser Ala
Arg His Pro Ala Trp Gly His Cys 420 425
430 Phe Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu
Pro Cys Thr 435 440 445
Ser Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450
455 460 Gly Thr Ser Arg
Trp Pro Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465 470
475 480 Thr Leu Gly Glu Gln Gly Leu Pro His
Gln Ser His Arg Gln Arg Ser 485 490
495 Ser Leu Ala Lys Ile Asn Ile Val Tyr Gln Glu Leu Asn Tyr
Arg Ser 500 505 510
Val Glu Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu Ser Ala Met
515 520 525 Gly Ser Leu Cys
Ser Leu Trp Phe Gly Ala Ser Val Leu Ser Leu Leu 530
535 540 Glu Leu Leu Glu Leu Leu Leu Asp
Ala Ser Ala Leu Thr Leu Val Leu 545 550
555 560 Gly Gly Arg Arg Leu Arg Arg Ala Trp Phe Ser Trp
Pro Arg Ala Ser 565 570
575 Pro Ala Ser Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln Met Pro
580 585 590 Pro Pro Ala
Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser Gly Pro His 595
600 605 Leu Pro Arg Val Met Leu Pro Gly
Val Leu Ala Gly Val Ser Ala Glu 610 615
620 Glu Ser Trp Ala Gly Pro Gln Pro Leu Glu Thr Leu Asp
Thr 625 630 635 19638PRTHomo
Sapiens 19Met Ala Glu His Arg Ser Met Asp Gly Arg Met Glu Ala Ala Thr Arg
1 5 10 15 Gly Gly
Pro His Leu Gln Ala Ala Ala Gln Thr Pro Pro Arg Pro Gly 20
25 30 Pro Pro Ser Ala Pro Pro Pro
Pro Pro Lys Glu Gly His Gln Glu Gly 35 40
45 Leu Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu
Thr Phe Phe Cys 50 55 60
Thr Asn Ala Thr Ile His Gly Ala Ile Arg Leu Val Cys Ser Arg Gly 65
70 75 80 Asn Arg Leu
Lys Thr Thr Ser Trp Gly Leu Leu Ser Leu Gly Ala Leu 85
90 95 Val Ala Leu Cys Trp Gln Leu Gly
Leu Leu Phe Glu Arg His Trp His 100 105
110 Arg Pro Val Leu Met Ala Val Ser Val His Ser Glu Arg
Lys Leu Leu 115 120 125
Pro Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro Ser Pro Val 130
135 140 Leu Arg His Leu
Glu Leu Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145 150
155 160 Ser Leu Tyr Asn Val Asn Leu Ser Lys
Gly Arg Ala Ala Leu Ser Ala 165 170
175 Thr Val Pro Pro His Glu Pro Pro Phe His Leu Asp Arg Glu
Ile Arg 180 185 190
Leu Gln Ser Leu Ser His Ser Gly Ser Arg Val Arg Val Gly Phe Arg
195 200 205 Leu Cys Asn Ser
Thr Gly Gly Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210
215 220 Gly Val Ala Ala Val Gln Asp Trp
Tyr His Phe His Tyr Val Asp Ile 225 230
235 240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp Ser His
Gly Ser Gln Asp 245 250
255 Gly His Phe Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln Ala
260 265 270 Arg Gln Phe
Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys Tyr Thr 275
280 285 Val Asp Gly Val Trp Thr Ala Gln
Arg Pro Gly Ile Thr His Gly Val 290 295
300 Gly Leu Val Leu Arg Val Glu Gln Gln Pro His Leu Pro
Leu Leu Ser 305 310 315
320 Thr Leu Ala Gly Ile Arg Val Met Val His Gly Arg Asn His Thr Pro
325 330 335 Phe Leu Gly His
His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala Thr 340
345 350 Ile Ser Ile Arg Glu Asp Glu Val His
Arg Leu Gly Ser Pro Tyr Gly 355 360
365 His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Glu Leu Leu
His Asn 370 375 380
Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser Cys Phe Gln Gln Leu 385
390 395 400 Met Val Glu Thr Cys
Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala 405
410 415 Gly Ala Glu Tyr Cys Ser Ser Ala Arg His
Pro Ala Trp Gly His Cys 420 425
430 Phe Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu Pro Cys
Thr 435 440 445 Ser
Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450
455 460 Gly Thr Ser Arg Trp Pro
Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465 470
475 480 Thr Leu Gly Glu Gln Gly Leu Pro His Gln Ser
His Arg Gln Arg Ser 485 490
495 Ser Leu Ala Lys Ile Asn Ile Val Tyr Gln Glu Leu Asn Tyr Arg Ser
500 505 510 Val Glu
Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu Ser Ala Met 515
520 525 Gly Ser Leu Cys Ser Leu Trp
Phe Gly Ala Ser Val Leu Ser Leu Leu 530 535
540 Glu Leu Leu Glu Leu Leu Leu Asp Ala Ser Ala Leu
Thr Leu Val Leu 545 550 555
560 Gly Gly Arg Arg Leu Arg Arg Ala Trp Phe Ser Trp Pro Arg Ala Ser
565 570 575 Pro Ala Ser
Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln Met Pro 580
585 590 Pro Pro Ala Gly Gly Thr Ser Asp
Asp Pro Glu Pro Ser Gly Pro His 595 600
605 Leu Pro Arg Val Met Leu Pro Gly Val Leu Ala Gly Val
Ser Ala Glu 610 615 620
Glu Ser Trp Ala Gly Pro Gln Pro Leu Glu Thr Leu Asp Lys 625
630 635 20638PRTHomo Sapiens 20Met Ala
Glu His Arg Ser Met Asp Gly Arg Met Glu Ala Ala Thr Arg 1 5
10 15 Gly Gly Pro His Leu Gln Ala
Ala Ala Gln Thr Pro Pro Arg Pro Gly 20 25
30 Pro Pro Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly
His Gln Glu Gly 35 40 45
Leu Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu Thr Phe Phe Cys
50 55 60 Thr Asn Ala
Thr Ile His Gly Ala Ile Arg Leu Val Cys Ser Arg Gly 65
70 75 80 Asn Arg Leu Lys Thr Thr Ser
Trp Gly Leu Leu Ser Leu Gly Ala Leu 85
90 95 Val Ala Leu Cys Trp Gln Leu Gly Leu Leu Phe
Glu Arg His Trp His 100 105
110 Arg Pro Val Leu Met Ala Val Ser Val His Ser Glu Arg Lys Leu
Leu 115 120 125 Pro
Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro Ser Pro Val 130
135 140 Leu Arg His Leu Glu Leu
Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145 150
155 160 Ser Leu Tyr Asn Val Asn Leu Ser Lys Gly Arg
Ala Ala Leu Ser Ala 165 170
175 Thr Val Pro Arg His Glu Pro Pro Phe His Leu Asp Arg Glu Ile Arg
180 185 190 Leu Gln
Arg Leu Ser His Ser Gly Ser Arg Val Arg Val Gly Phe Arg 195
200 205 Leu Cys Asn Ser Thr Gly Gly
Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210 215
220 Gly Val Ala Ala Val Gln Asp Trp Tyr His Phe His
Tyr Val Asp Ile 225 230 235
240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp Ser His Gly Ser Gln Asp
245 250 255 Gly His Phe
Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln Ala 260
265 270 Arg Gln Phe Arg Thr Phe His His
Pro Thr Tyr Gly Ser Cys Tyr Thr 275 280
285 Val Asp Gly Val Trp Thr Ala Gln Arg Pro Gly Ile Thr
His Gly Val 290 295 300
Gly Leu Val Leu Arg Val Glu Gln Gln Pro His Leu Pro Leu Leu Ser 305
310 315 320 Thr Leu Ala Gly
Ile Arg Val Met Val His Gly Arg Asn His Thr Pro 325
330 335 Phe Leu Gly His His Ser Phe Ser Val
Arg Pro Gly Thr Glu Ala Thr 340 345
350 Ile Ser Ile Arg Glu Asp Glu Val His Arg Leu Gly Ser Pro
Tyr Gly 355 360 365
His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Glu Leu Leu His Asn 370
375 380 Thr Ser Tyr Thr Arg
Gln Ala Cys Leu Val Ser Cys Phe Gln Gln Leu 385 390
395 400 Met Val Glu Thr Cys Ser Cys Gly Tyr Tyr
Leu His Pro Leu Pro Ala 405 410
415 Gly Ala Glu Tyr Cys Ser Ser Ala Arg His Pro Ala Trp Gly His
Cys 420 425 430 Phe
Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu Pro Cys Thr 435
440 445 Ser Arg Cys Pro Arg Pro
Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450 455
460 Gly Thr Ser Arg Trp Pro Ser Ala Lys Ser Ala
Gly Trp Thr Leu Ala 465 470 475
480 Thr Leu Gly Glu Gln Gly Leu Pro His Gln Ser His Arg Gln Arg Ser
485 490 495 Ser Leu
Ala Lys Ile Asn Ile Val Tyr Gln Gly Leu Asn Tyr Arg Ser 500
505 510 Val Glu Glu Ala Pro Val Tyr
Ser Val Pro Gln Leu Leu Ser Ala Met 515 520
525 Gly Ser Leu Cys Ser Leu Trp Phe Gly Ala Ser Val
Leu Ser Leu Leu 530 535 540
Glu Leu Leu Glu Leu Leu Leu Asp Ala Ser Ala Leu Thr Leu Val Leu 545
550 555 560 Gly Gly Arg
Arg Leu Arg Arg Ala Trp Phe Ser Trp Pro Arg Ala Ser 565
570 575 Pro Ala Ser Gly Ala Ser Ser Ile
Lys Pro Glu Ala Ser Gln Met Pro 580 585
590 Pro Pro Ala Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser
Gly Pro His 595 600 605
Leu Pro Arg Val Met Leu Pro Gly Val Leu Ala Gly Val Ser Ala Glu 610
615 620 Glu Ser Trp Ala
Gly Pro Gln Pro Leu Glu Thr Leu Asp Thr 625 630
635 21638PRTHomo Sapiens 21Met Ala Glu His Arg Ser Met
Asp Gly Arg Met Glu Ala Ala Thr Arg 1 5
10 15 Gly Gly Ser His Leu Gln Ala Ala Ala Gln Thr
Pro Pro Arg Pro Gly 20 25
30 Pro Pro Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly His Gln Glu
Gly 35 40 45 Leu
Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu Thr Phe Phe Cys 50
55 60 Thr Asn Ala Thr Ile His
Gly Thr Ile Arg Leu Val Cys Ser Arg Gly 65 70
75 80 Asn Arg Leu Lys Thr Thr Ser Trp Gly Leu Leu
Ser Leu Gly Ala Leu 85 90
95 Val Ala Leu Cys Trp Gln Leu Gly Leu Leu Phe Glu Arg His Trp His
100 105 110 Arg Pro
Val Leu Met Ala Val Ser Val His Ser Glu Arg Lys Leu Leu 115
120 125 Pro Leu Val Thr Leu Cys Asp
Gly Asn Pro Arg Arg Pro Ser Pro Val 130 135
140 Leu Arg His Leu Glu Leu Leu Asp Glu Phe Ala Arg
Glu Asn Ile Asp 145 150 155
160 Ser Leu Tyr Asn Val Asn Leu Ser Lys Gly Arg Ala Ala Leu Ser Ala
165 170 175 Thr Val Pro
Pro His Glu Pro Pro Phe His Leu Asp Arg Glu Ile Arg 180
185 190 Leu Gln Arg Leu Ser His Ser Gly
Ser Arg Val Arg Val Gly Phe Arg 195 200
205 Leu Cys Asn Ser Thr Gly Gly Asp Cys Phe Tyr Arg Gly
Tyr Thr Ser 210 215 220
Gly Val Ala Ala Val Gln Asp Trp Tyr His Phe His Tyr Val Asp Ile 225
230 235 240 Leu Ala Leu Leu
Pro Ala Ala Trp Glu Asp Ser His Gly Ser Gln Asp 245
250 255 Gly His Phe Val Leu Ser Cys Ser Tyr
Asp Gly Leu Asp Cys Gln Ala 260 265
270 Arg Gln Phe Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys
Tyr Thr 275 280 285
Val Asp Gly Val Trp Thr Ala Gln Arg Pro Gly Ile Thr His Gly Val 290
295 300 Gly Leu Val Leu Arg
Val Glu Gln Gln Pro His Leu Pro Leu Leu Ser 305 310
315 320 Thr Leu Ala Gly Ile Arg Val Met Val His
Gly Arg Asn His Thr Pro 325 330
335 Phe Leu Gly His His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala
Thr 340 345 350 Ile
Ser Ile Arg Glu Asp Glu Val His Arg Leu Gly Ser Pro Tyr Gly 355
360 365 His Cys Thr Ala Gly Gly
Glu Gly Val Glu Val Gln Leu Leu His Asn 370 375
380 Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser
Cys Phe Gln Gln Leu 385 390 395
400 Met Val Glu Thr Cys Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala
405 410 415 Gly Ala
Glu Tyr Cys Ser Ser Ala Arg His Pro Ala Trp Gly His Cys 420
425 430 Phe Tyr Arg Leu Tyr Arg Asp
Leu Glu Thr His Arg Leu Pro Cys Thr 435 440
445 Ser Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe
Lys Leu Ser Thr 450 455 460
Gly Thr Ser Arg Trp Pro Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465
470 475 480 Thr Leu Gly
Glu Gln Gly Leu Pro His Gln Ser His Arg Gln Arg Ser 485
490 495 Ser Leu Ala Lys Ile Asn Ile Val
Tyr Gln Glu Leu Asn Tyr Arg Ser 500 505
510 Val Glu Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu
Ser Ala Met 515 520 525
Gly Ser Leu Cys Ser Leu Trp Phe Gly Ala Ser Val Leu Ser Leu Leu 530
535 540 Glu Leu Leu Glu
Leu Leu Leu Asp Ala Ser Ala Leu Thr Leu Val Leu 545 550
555 560 Gly Gly Arg Arg Leu Arg Arg Ala Trp
Phe Ser Trp Pro Arg Ala Ser 565 570
575 Pro Ala Ser Gly Ala Ser Ser Ile Lys Pro Glu Ala Gly Gln
Met Pro 580 585 590
Pro Pro Ala Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser Gly Pro His
595 600 605 Leu Pro Arg Val
Met Leu Pro Gly Val Leu Ala Gly Val Ser Ala Glu 610
615 620 Glu Ser Trp Ala Gly Pro Gln Pro
Leu Glu Thr Leu Asp Thr 625 630 635
22638PRTHomo Sapiens 22Met Ala Glu His Arg Ser Met Asp Gly Arg Met
Glu Ala Ala Thr Arg 1 5 10
15 Gly Gly Ser His Leu Gln Ala Ala Ala Gln Thr Pro Pro Arg Pro Gly
20 25 30 Pro Pro
Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly His Gln Glu Gly 35
40 45 Leu Val Glu Leu Pro Ala Ser
Phe Arg Glu Leu Leu Thr Phe Phe Cys 50 55
60 Thr Asn Ala Thr Ile His Gly Ala Ile Arg Leu Val
Cys Ser Arg Gly 65 70 75
80 Asn Arg Leu Lys Thr Thr Ser Trp Gly Leu Leu Ser Leu Gly Ala Leu
85 90 95 Val Ala Leu
Cys Trp Gln Leu Gly Leu Leu Phe Glu Arg His Trp His 100
105 110 Arg Pro Val Leu Met Ala Val Ser
Val His Ser Glu Arg Lys Leu Leu 115 120
125 Pro Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro
Ser Pro Val 130 135 140
Leu Arg His Leu Glu Leu Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145
150 155 160 Ser Leu Tyr Asn
Val Asn Leu Ser Lys Gly Arg Ala Ala Leu Ser Ala 165
170 175 Thr Val Pro Arg His Glu Pro Pro Phe
His Leu Asp Arg Glu Ile Arg 180 185
190 Leu Gln Arg Leu Ser His Ser Gly Ser Arg Val Arg Val Gly
Phe Arg 195 200 205
Leu Cys Asn Ser Thr Gly Gly Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210
215 220 Gly Val Ala Ala Val
Gln Asp Trp Tyr His Phe His Tyr Val Asp Ile 225 230
235 240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp
Ser His Gly Ser Gln Asp 245 250
255 Gly His Phe Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln
Ala 260 265 270 Arg
Gln Phe Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys Tyr Thr 275
280 285 Val Asp Gly Val Trp Thr
Ala Gln Arg Pro Gly Ile Thr His Gly Val 290 295
300 Gly Leu Val Leu Arg Val Glu Gln Gln Pro His
Leu Pro Leu Leu Ser 305 310 315
320 Thr Leu Ala Gly Ile Arg Val Met Val His Gly Arg Asn His Thr Pro
325 330 335 Phe Leu
Gly His His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala Thr 340
345 350 Ile Ser Ile Arg Glu Asp Glu
Val His Arg Leu Gly Ser Pro Tyr Gly 355 360
365 His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Glu
Leu Leu His Asn 370 375 380
Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser Cys Phe Gln Gln Leu 385
390 395 400 Met Val Glu
Thr Cys Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala 405
410 415 Gly Ala Glu Tyr Cys Ser Ser Ala
Arg His Pro Ala Trp Gly His Cys 420 425
430 Phe Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu
Pro Cys Thr 435 440 445
Ser Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450
455 460 Gly Thr Ser Arg
Trp Pro Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465 470
475 480 Thr Leu Gly Glu Gln Gly Leu Pro His
Gln Gly His Arg Gln Arg Ser 485 490
495 Ser Leu Ala Lys Ile Asn Ile Val Tyr Gln Glu Leu Asn Tyr
Arg Ser 500 505 510
Val Glu Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu Ser Ala Met
515 520 525 Gly Ser Leu Cys
Ser Leu Trp Phe Gly Ala Ser Val Leu Ser Leu Leu 530
535 540 Glu Leu Leu Glu Leu Leu Leu Asp
Ala Ser Ala Leu Thr Leu Val Leu 545 550
555 560 Gly Gly Arg Arg Leu Arg Arg Ala Trp Phe Ser Trp
Pro Arg Ala Ser 565 570
575 Pro Ala Ser Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln Met Pro
580 585 590 Pro Pro Ala
Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser Gly Pro His 595
600 605 Leu Pro Arg Val Met Leu Pro Gly
Val Leu Ala Gly Val Ser Ala Glu 610 615
620 Glu Ser Trp Ala Gly Pro Gln Pro Leu Glu Thr Leu Asp
Thr 625 630 635 23638PRTHomo
Sapiens 23Met Ala Glu His Arg Ser Met Asp Gly Arg Met Glu Ala Ala Thr Arg
1 5 10 15 Gly Gly
Ser His Leu Gln Ala Ala Ala Gln Thr Pro Pro Arg Pro Gly 20
25 30 Pro Pro Ser Ala Pro Pro Pro
Pro Pro Lys Glu Gly His Gln Glu Gly 35 40
45 Leu Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu
Thr Phe Phe Cys 50 55 60
Thr Asn Ala Thr Ile His Gly Ala Ile Arg Leu Val Cys Ser Arg Gly 65
70 75 80 Asn Arg Leu
Lys Thr Thr Ser Trp Gly Leu Leu Ser Leu Gly Ala Leu 85
90 95 Val Ala Leu Cys Trp Gln Leu Gly
Leu Leu Phe Glu Arg His Trp His 100 105
110 Arg Pro Val Leu Met Ala Val Ser Val His Ser Glu Arg
Lys Leu Leu 115 120 125
Pro Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro Ser Pro Val 130
135 140 Leu Arg His Leu
Glu Leu Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145 150
155 160 Ser Leu Tyr Asn Val Asn Leu Ser Lys
Gly Arg Ala Ala Leu Ser Ala 165 170
175 Thr Val Pro Pro His Glu Pro Pro Phe His Leu Asp Arg Glu
Ile Arg 180 185 190
Leu Gln Arg Leu Ser His Ser Gly Ser Arg Val Arg Val Gly Phe Arg
195 200 205 Leu Cys Asn Ser
Thr Gly Gly Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210
215 220 Gly Val Ala Ala Val Gln Asp Trp
Tyr His Phe His Tyr Val Asp Ile 225 230
235 240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp Ser His
Gly Ser Gln Asp 245 250
255 Gly His Phe Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln Ala
260 265 270 Arg Gln Phe
Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys Tyr Thr 275
280 285 Val Asp Gly Val Trp Thr Ala Gln
Arg Pro Gly Ile Thr His Gly Val 290 295
300 Gly Leu Val Leu Arg Val Glu Gln Gln Pro His Leu Pro
Leu Leu Ser 305 310 315
320 Thr Leu Ala Gly Ile Arg Val Met Val His Gly Arg Asn His Thr Pro
325 330 335 Phe Leu Gly His
His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala Thr 340
345 350 Ile Ser Ile Arg Glu Asp Glu Val His
Arg Leu Gly Ser Pro Tyr Gly 355 360
365 His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Gln Pro Leu
His Asn 370 375 380
Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser Cys Phe Gln Gln Leu 385
390 395 400 Met Val Glu Thr Cys
Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala 405
410 415 Gly Ala Glu Tyr Cys Ser Ser Ala Arg His
Pro Ala Trp Gly His Cys 420 425
430 Phe Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu Pro Cys
Thr 435 440 445 Ser
Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450
455 460 Gly Thr Ser Arg Trp Pro
Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465 470
475 480 Thr Leu Gly Glu Gln Gly Leu Pro His Gln Ser
His Arg Gln Arg Ser 485 490
495 Ser Leu Ala Lys Ile Asn Ile Val Tyr Gln Glu Leu Asn Tyr Arg Ser
500 505 510 Val Glu
Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu Ser Ala Met 515
520 525 Gly Ser Leu Cys Ser Leu Trp
Phe Gly Ala Ser Val Leu Ser Leu Leu 530 535
540 Glu Leu Leu Glu Leu Leu Leu Asp Ala Ser Ala Leu
Thr Leu Val Leu 545 550 555
560 Gly Gly Arg Arg Leu Arg Arg Ala Trp Phe Ser Trp Pro Arg Ala Ser
565 570 575 Pro Ala Ser
Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln Met Pro 580
585 590 Pro Pro Ala Gly Gly Thr Ser Asp
Asp Pro Glu Pro Ser Gly Pro His 595 600
605 Leu Pro Arg Val Met Leu Pro Gly Val Leu Ala Gly Val
Ser Ala Glu 610 615 620
Glu Ser Trp Ala Gly Pro Gln Pro Leu Glu Thr Leu Asp Thr 625
630 635 24638PRTHomo Sapiens 24Met Ala
Glu His Arg Ser Met Asp Gly Arg Met Glu Ala Ala Thr Arg 1 5
10 15 Gly Gly Ser His Leu Gln Ala
Ala Ala Gln Thr Pro Pro Arg Pro Gly 20 25
30 Pro Pro Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly
His Gln Glu Gly 35 40 45
Leu Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu Thr Phe Phe Cys
50 55 60 Thr Asn Ala
Thr Ile His Gly Ala Ile Arg Leu Val Cys Ser Arg Gly 65
70 75 80 Asn Arg Leu Lys Thr Thr Ser
Trp Gly Leu Leu Ser Leu Gly Ala Leu 85
90 95 Val Ala Leu Cys Trp Gln Leu Gly Leu Leu Phe
Glu Arg His Trp His 100 105
110 Arg Pro Val Leu Met Ala Val Ser Val His Ser Glu Arg Lys Leu
Leu 115 120 125 Pro
Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro Ser Pro Val 130
135 140 Leu Arg His Leu Glu Leu
Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145 150
155 160 Ser Leu Tyr Ser Val Asn Leu Ser Lys Gly Arg
Ala Ala Leu Ser Ala 165 170
175 Thr Val Pro Arg His Glu Pro Pro Phe His Leu Asp Arg Glu Ile Arg
180 185 190 Leu Gln
Arg Leu Ser His Ser Gly Ser Arg Val Arg Val Gly Phe Arg 195
200 205 Leu Cys Asn Ser Thr Gly Gly
Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210 215
220 Gly Val Ala Ala Val Gln Asp Trp Tyr His Phe His
Tyr Val Asp Ile 225 230 235
240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp Ser His Gly Ser Gln Asp
245 250 255 Gly His Phe
Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln Ala 260
265 270 Arg Gln Phe Arg Thr Phe His His
Pro Thr Tyr Gly Ser Cys Tyr Thr 275 280
285 Val Asp Gly Val Trp Thr Ala Gln Arg Pro Gly Ile Thr
His Gly Val 290 295 300
Gly Leu Val Leu Arg Val Glu Gln Gln Pro His Leu Pro Leu Leu Ser 305
310 315 320 Thr Leu Ala Gly
Ile Arg Val Met Val His Gly Arg Asn His Thr Pro 325
330 335 Phe Leu Gly His His Ser Phe Ser Val
Arg Pro Gly Thr Glu Ala Thr 340 345
350 Ile Ser Ile Arg Glu Asp Glu Val His Arg Leu Gly Ser Pro
Tyr Gly 355 360 365
His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Glu Leu Leu His Asn 370
375 380 Thr Ser Tyr Thr Arg
Gln Pro Cys Leu Val Ser Cys Phe Gln Gln Leu 385 390
395 400 Met Val Glu Thr Cys Ser Cys Gly Tyr Tyr
Leu His Pro Leu Pro Ala 405 410
415 Gly Ala Glu Tyr Cys Ser Ser Ala Arg His Pro Ala Trp Gly His
Cys 420 425 430 Phe
Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu Pro Cys Thr 435
440 445 Ser Arg Cys Pro Arg Pro
Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450 455
460 Gly Thr Ser Arg Trp Pro Ser Ala Lys Ser Ala
Gly Trp Thr Leu Ala 465 470 475
480 Thr Leu Gly Glu Gln Gly Leu Pro His Gln Ser His Arg Gln Arg Ser
485 490 495 Ser Leu
Ala Lys Ile Asn Ile Val Tyr Gln Glu Leu Asn Tyr Arg Ser 500
505 510 Val Glu Glu Ala Pro Val Tyr
Ser Val Pro Gln Leu Leu Ser Ala Met 515 520
525 Gly Ser Leu Cys Ser Leu Trp Phe Gly Ala Ser Val
Leu Ser Leu Leu 530 535 540
Glu Leu Leu Glu Leu Leu Leu Asp Ala Ser Ala Leu Thr Leu Val Leu 545
550 555 560 Gly Gly Arg
Arg Leu Arg Arg Ala Trp Phe Ser Trp Pro Arg Ala Ser 565
570 575 Pro Ala Ser Gly Ala Ser Ser Ile
Lys Pro Glu Ala Ser Gln Met Pro 580 585
590 Pro Pro Ala Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser
Gly Pro His 595 600 605
Leu Pro Arg Val Met Leu Pro Gly Val Leu Ala Gly Val Ser Ala Glu 610
615 620 Glu Ser Trp Ala
Gly Pro Gln Pro Leu Glu Thr Leu Asp Thr 625 630
635 25638PRTHomo Sapiens 25Met Ala Glu His Arg Ser Met
Asp Gly Arg Met Glu Ala Ala Thr Arg 1 5
10 15 Gly Gly Ser His Leu Gln Ala Ala Ala Gln Thr
Pro Pro Arg Pro Gly 20 25
30 Pro Pro Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly His Gln Glu
Gly 35 40 45 Leu
Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu Thr Phe Phe Cys 50
55 60 Thr Asn Ala Thr Ile His
Gly Ala Ile Arg Leu Val Cys Ser Arg Gly 65 70
75 80 Asn Arg Leu Lys Thr Thr Ser Trp Gly Leu Leu
Ser Leu Gly Ala Leu 85 90
95 Val Ala Leu Cys Trp Gln Leu Gly Leu Leu Phe Glu Arg His Trp His
100 105 110 Arg Pro
Val Leu Met Ala Val Ser Val His Ser Glu Arg Lys Leu Leu 115
120 125 Pro Leu Val Thr Leu Cys Asp
Gly Asn Pro Arg Arg Pro Ser Pro Val 130 135
140 Leu Arg His Leu Glu Leu Leu Asp Glu Phe Ala Arg
Glu Asn Ile Asp 145 150 155
160 Ser Leu Tyr Asn Val Asn Leu Ser Lys Gly Arg Ala Ala Leu Ser Ala
165 170 175 Thr Val Pro
Arg His Glu Pro Pro Phe His Leu Asp Arg Glu Ile Arg 180
185 190 Leu Gln Arg Leu Ser His Ser Gly
Ser Arg Val Arg Val Gly Phe Arg 195 200
205 Leu Cys Asn Ser Thr Gly Gly Asp Cys Phe Tyr Arg Gly
Tyr Thr Ser 210 215 220
Gly Val Ala Ala Val Gln Asp Trp Tyr His Phe His Tyr Val Asp Ile 225
230 235 240 Leu Ala Leu Leu
Pro Ala Ala Trp Glu Asp Ser His Gly Ser Gln Asp 245
250 255 Gly His Phe Val Leu Ser Cys Ser Tyr
Asp Gly Leu Asp Cys Gln Ala 260 265
270 Arg Gln Phe Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys
Tyr Thr 275 280 285
Val Asp Gly Val Trp Thr Ala Gln Arg Pro Gly Ile Thr His Gly Val 290
295 300 Gly Leu Val Leu Arg
Val Glu Gln Gln Pro His Leu Pro Leu Leu Ser 305 310
315 320 Thr Leu Ala Gly Ile Arg Val Met Val His
Gly Arg Asn His Thr Pro 325 330
335 Phe Leu Gly His His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala
Thr 340 345 350 Ile
Ser Ile Arg Glu Asp Glu Val His Arg Leu Gly Ser Pro Tyr Gly 355
360 365 His Cys Thr Ala Gly Gly
Glu Gly Val Glu Val Glu Leu Leu His Asn 370 375
380 Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser
Cys Phe Gln Gln Leu 385 390 395
400 Met Val Glu Thr Cys Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala
405 410 415 Gly Ala
Glu Tyr Cys Gly Ser Ala Arg His Pro Ala Trp Gly His Cys 420
425 430 Phe Tyr Arg Leu Tyr Gln Asp
Leu Glu Thr His Arg Leu Pro Cys Thr 435 440
445 Ser Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe
Lys Leu Ser Thr 450 455 460
Gly Thr Ser Arg Trp Pro Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465
470 475 480 Thr Leu Gly
Glu Gln Gly Leu Pro His Gln Ser His Arg Gln Arg Ser 485
490 495 Ser Leu Ala Lys Ile Asn Ile Val
Tyr Gln Glu Leu Asn Tyr Arg Ser 500 505
510 Val Glu Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu
Ser Ala Met 515 520 525
Gly Ser Leu Cys Ser Leu Trp Phe Gly Ala Ser Val Leu Ser Leu Leu 530
535 540 Glu Leu Leu Glu
Leu Leu Leu Asp Ala Ser Ala Leu Thr Leu Val Leu 545 550
555 560 Gly Gly Arg Arg Leu Arg Arg Ala Trp
Phe Ser Trp Pro Arg Ala Ser 565 570
575 Pro Ala Ser Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln
Met Pro 580 585 590
Pro Pro Ala Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser Gly Pro His
595 600 605 Leu Pro Arg Val
Met Leu Pro Gly Val Leu Ala Gly Val Ser Ala Glu 610
615 620 Glu Ser Trp Ala Gly Pro Gln Pro
Leu Glu Thr Leu Asp Thr 625 630 635
26638PRTHomo Sapiens 26Met Ala Glu His Arg Ser Met Asp Gly Arg Met
Glu Ala Ala Thr Arg 1 5 10
15 Gly Gly Ser His Leu Gln Ala Ala Ala Gln Thr Pro Pro Arg Pro Gly
20 25 30 Pro Pro
Ser Ala Pro Pro Pro Pro Pro Lys Glu Gly His Gln Glu Gly 35
40 45 Leu Val Glu Leu Pro Ala Ser
Phe Arg Glu Leu Leu Thr Phe Phe Cys 50 55
60 Thr Asn Ala Thr Ile His Gly Ala Ile Arg Leu Val
Cys Ser Arg Gly 65 70 75
80 Asn Arg Leu Lys Thr Thr Ser Trp Gly Leu Leu Ser Leu Gly Ala Leu
85 90 95 Val Ala Leu
Cys Trp Gln Leu Gly Leu Leu Phe Glu Arg His Trp His 100
105 110 Arg Pro Val Leu Met Ala Val Ser
Val His Ser Glu Arg Lys Leu Leu 115 120
125 Pro Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro
Ser Pro Val 130 135 140
Leu Arg His Leu Glu Leu Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145
150 155 160 Ser Leu Tyr Asn
Val Asn Leu Ser Lys Gly Arg Ala Ala Leu Ser Ala 165
170 175 Thr Val Pro Arg His Glu Pro Pro Phe
His Leu Asp Arg Glu Ile Arg 180 185
190 Leu Gln Arg Leu Ser His Ser Gly Ser Arg Val Arg Val Gly
Phe Arg 195 200 205
Leu Cys Asn Ser Thr Gly Gly Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210
215 220 Gly Val Ala Ala Val
Gln Asp Trp Tyr His Phe His Tyr Val Asp Ile 225 230
235 240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp
Ser His Gly Ser Gln Asp 245 250
255 Gly His Phe Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln
Ala 260 265 270 Arg
Gln Phe Arg Thr Ile His His Pro Thr Tyr Gly Ser Cys Tyr Thr 275
280 285 Val Asp Gly Val Trp Thr
Ala Gln Arg Pro Gly Ile Thr His Gly Val 290 295
300 Gly Leu Val Leu Arg Val Glu Gln Gln Pro His
Leu Pro Leu Leu Ser 305 310 315
320 Thr Leu Ala Gly Ile Arg Val Met Val His Gly Arg Asn His Thr Pro
325 330 335 Phe Leu
Gly His His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala Thr 340
345 350 Ile Arg Ile Arg Glu Asp Glu
Val His Arg Leu Gly Ser Pro Tyr Gly 355 360
365 His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Glu
Leu Leu His Asn 370 375 380
Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser Cys Phe Gln Gln Leu 385
390 395 400 Met Val Glu
Thr Cys Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala 405
410 415 Gly Ala Glu Tyr Cys Ser Ser Ala
Arg His Pro Ala Trp Gly His Cys 420 425
430 Phe Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu
Pro Cys Thr 435 440 445
Ser Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450
455 460 Gly Thr Ser Arg
Trp Pro Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465 470
475 480 Thr Leu Gly Glu Gln Gly Leu Pro His
Gln Ser His Arg Gln Arg Ser 485 490
495 Ser Leu Ala Lys Ile Asn Ile Val Tyr Gln Glu Leu Asn Tyr
Arg Ser 500 505 510
Val Glu Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu Ser Ala Met
515 520 525 Gly Ser Leu Cys
Ser Leu Trp Phe Gly Ala Ser Val Leu Ser Leu Leu 530
535 540 Glu Leu Leu Glu Leu Leu Leu Asp
Ala Ser Ala Leu Thr Leu Val Leu 545 550
555 560 Gly Gly Arg Arg Leu His Arg Ala Trp Phe Ser Trp
Pro Arg Ala Ser 565 570
575 Pro Ala Ser Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln Met Pro
580 585 590 Pro Pro Ala
Gly Gly Thr Ser Asp Asp Pro Glu Pro Ser Gly Pro His 595
600 605 Leu Pro Arg Val Met Leu Pro Gly
Val Leu Ala Gly Val Ser Ala Glu 610 615
620 Glu Ser Trp Ala Gly Pro Gln Pro Leu Glu Thr Leu Asp
Thr 625 630 635 27649PRTHomo
Sapiens 27Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 28649PRTHomo
Sapiens 28Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val Asn Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 29649PRTHomo
Sapiens 29Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 30649PRTHomo
Sapiens 30Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Glu Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Ser
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 31649PRTHomo
Sapiens 31Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Ala 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Thr Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Ser Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 32649PRTHomo
Sapiens 32Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Gly Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 33649PRTHomo
Sapiens 33Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Pro 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Leu Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Leu Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Thr Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 34649PRTHomo
Sapiens 34Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 35649PRTHomo
Sapiens 35Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 36649PRTHomo
Sapiens 36Met Ala Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro
1 5 10 15 Val Thr
Gly Pro Gln Ala Pro Thr Ile Lys Glu Leu Met Arg Trp Tyr 20
25 30 Cys Leu Asn Thr Asn Thr His
Gly Cys Arg Arg Ile Val Val Ser Arg 35 40
45 Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr
Leu Thr Ala Val 50 55 60
Ala Leu Ile Leu Arg Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val
Ser Ile Lys Val His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro
Tyr Lys Tyr Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu
Lys Ser Leu 115 120 125
Tyr Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu
Gly Lys Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly
Lys Ala Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala
Ser Asn 180 185 190
Val Met His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser
195 200 205 Asn Asp Thr Ser
Asp Cys Ala Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210
215 220 Ala Ile Gln Glu Trp Tyr Lys Leu
His Tyr Met Asn Ile Met Ala Gln 225 230
235 240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser
Ala Glu Glu Leu 245 250
255 Leu Val Thr Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe
260 265 270 Thr Leu Phe
His His Pro Met His Gly Asn Cys Tyr Thr Phe Asn Asn 275
280 285 Arg Glu Asn Glu Thr Ile Leu Ser
Thr Ser Met Gly Gly Ser Glu Tyr 290 295
300 Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr
Asn Pro Phe 305 310 315
320 Leu Val Ser Ser Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu
325 330 335 Tyr Pro Phe Val
Glu Asp Val Gly Thr Glu Ile Glu Thr Ala Met Val 340
345 350 Thr Ser Ile Gly Met His Leu Thr Glu
Ser Phe Lys Leu Ser Glu Pro 355 360
365 Tyr Ser Gln Cys Thr Glu Asp Gly Ser Asp Val Pro Ile Arg
Asn Ile 370 375 380
Tyr Asn Ala Ala Tyr Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385
390 395 400 Thr Lys Met Val Glu
Lys Cys Gly Cys Ala Gln Tyr Ser Gln Pro Leu 405
410 415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln
Gln His Pro Asn Trp Met 420 425
430 Tyr Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu
Gly 435 440 445 Cys
Gln Ser Val Cys Lys Glu Ala Cys Ser Phe Lys Glu Trp Thr Leu 450
455 460 Thr Thr Ser Leu Ala Gln
Trp Pro Ser Val Val Ser Glu Lys Trp Leu 465 470
475 480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln
Val Asn Lys Lys Leu 485 490
495 Asn Lys Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn
500 505 510 Gln Arg
Ser Ile Met Glu Ser Pro Ala Asn Ser Ile Glu Met Leu Leu 515
520 525 Ser Asn Phe Gly Gly Gln Leu
Gly Leu Trp Met Ser Cys Ser Val Val 530 535
540 Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp
Phe Phe Ser Ile 545 550 555
560 Ile Ala Arg Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys
565 570 575 Gln Ala Pro
Pro Cys Pro Glu Ala Pro Arg Ser Pro Gln Gly Gln Asp 580
585 590 Asn Pro Ala Leu Asp Ile Asp Asp
Asp Leu Pro Thr Phe Asn Ser Ala 595 600
605 Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly
Thr Pro Pro 610 615 620
Pro Lys Tyr Asn Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625
630 635 640 Thr Asp Thr Gln
Met Leu Asp Glu Leu 645 37638PRTHomo
sapiens 37Met Ala Glu His Arg Ser Met Asp Gly Arg Met Glu Ala Ala Thr Arg
1 5 10 15 Gly Gly
Ser His Leu Gln Ala Ala Ala Gln Thr Pro Pro Arg Pro Gly 20
25 30 Pro Pro Ser Ala Pro Pro Pro
Pro Pro Lys Glu Gly His Gln Glu Gly 35 40
45 Leu Val Glu Leu Pro Ala Ser Phe Arg Glu Leu Leu
Thr Phe Phe Cys 50 55 60
Thr Asn Ala Thr Ile His Gly Ala Ile Arg Leu Val Cys Ser Arg Gly 65
70 75 80 Asn Arg Leu
Lys Thr Thr Ser Trp Gly Leu Leu Ser Leu Gly Ala Leu 85
90 95 Val Ala Leu Cys Trp Gln Leu Gly
Leu Leu Phe Glu Arg His Trp His 100 105
110 Arg Pro Val Leu Met Ala Val Ser Val His Ser Glu Arg
Lys Leu Leu 115 120 125
Pro Leu Val Thr Leu Cys Asp Gly Asn Pro Arg Arg Pro Ser Pro Val 130
135 140 Leu Arg His Leu
Glu Leu Leu Asp Glu Phe Ala Arg Glu Asn Ile Asp 145 150
155 160 Ser Leu Tyr Asn Val Asn Leu Ser Lys
Gly Arg Ala Ala Leu Ser Ala 165 170
175 Thr Val Pro Arg His Glu Pro Pro Phe His Leu Asp Arg Glu
Ile Arg 180 185 190
Leu Gln Arg Leu Ser His Ser Gly Ser Arg Val Arg Val Gly Phe Arg
195 200 205 Leu Cys Asn Ser
Thr Gly Gly Asp Cys Phe Tyr Arg Gly Tyr Thr Ser 210
215 220 Gly Val Ala Ala Val Gln Asp Trp
Tyr His Phe His Tyr Val Asp Ile 225 230
235 240 Leu Ala Leu Leu Pro Ala Ala Trp Glu Asp Ser His
Gly Ser Gln Asp 245 250
255 Gly His Phe Val Leu Ser Cys Ser Tyr Asp Gly Leu Asp Cys Gln Ala
260 265 270 Arg Gln Phe
Arg Thr Phe His His Pro Thr Tyr Gly Ser Cys Tyr Thr 275
280 285 Val Asp Gly Val Trp Thr Ala Gln
Arg Pro Gly Ile Thr His Gly Val 290 295
300 Gly Leu Val Leu Arg Val Glu Gln Gln Pro His Leu Pro
Leu Leu Ser 305 310 315
320 Thr Leu Ala Gly Ile Arg Val Met Val His Gly Arg Asn His Thr Pro
325 330 335 Phe Leu Gly His
His Ser Phe Ser Val Arg Pro Gly Thr Glu Ala Thr 340
345 350 Ile Ser Ile Arg Glu Asp Glu Val His
Arg Leu Gly Ser Pro Tyr Gly 355 360
365 His Cys Thr Ala Gly Gly Glu Gly Val Glu Val Glu Leu Leu
His Asn 370 375 380
Thr Ser Tyr Thr Arg Gln Ala Cys Leu Val Ser Cys Phe Gln Gln Leu 385
390 395 400 Met Val Glu Thr Cys
Ser Cys Gly Tyr Tyr Leu His Pro Leu Pro Ala 405
410 415 Gly Ala Glu Tyr Cys Ser Ser Ala Arg His
Pro Ala Trp Gly His Cys 420 425
430 Phe Tyr Arg Leu Tyr Gln Asp Leu Glu Thr His Arg Leu Pro Cys
Thr 435 440 445 Ser
Arg Cys Pro Arg Pro Cys Arg Glu Ser Ala Phe Lys Leu Ser Thr 450
455 460 Gly Thr Ser Arg Trp Pro
Ser Ala Lys Ser Ala Gly Trp Thr Leu Ala 465 470
475 480 Thr Leu Gly Glu Gln Gly Leu Pro His Gln Ser
His Arg Gln Arg Ser 485 490
495 Ser Leu Ala Lys Ile Asn Ile Val Tyr Gln Glu Leu Asn Tyr Arg Ser
500 505 510 Val Glu
Glu Ala Pro Val Tyr Ser Val Pro Gln Leu Leu Ser Ala Met 515
520 525 Gly Ser Leu Tyr Ser Leu Trp
Phe Gly Ala Ser Val Leu Ser Leu Leu 530 535
540 Glu Leu Leu Glu Leu Leu Leu Asp Ala Ser Ala Leu
Thr Leu Val Leu 545 550 555
560 Gly Gly Arg Arg Leu Arg Arg Ala Trp Phe Ser Trp Pro Arg Ala Ser
565 570 575 Pro Ala Ser
Gly Ala Ser Ser Ile Lys Pro Glu Ala Ser Gln Met Pro 580
585 590 Pro Pro Ala Gly Gly Thr Ser Asp
Asp Pro Glu Pro Ser Gly Pro His 595 600
605 Leu Pro Arg Val Met Leu Pro Gly Val Leu Ala Gly Val
Ser Ala Glu 610 615 620
Glu Ser Trp Ala Gly Pro Gln Pro Leu Glu Thr Leu Asp Thr 625
630 635 38649PRTHomo sapiens 38Met Ala
Pro Gly Glu Lys Ile Lys Ala Lys Ile Lys Lys Asn Leu Pro 1 5
10 15 Val Thr Gly Pro Gln Ala Pro
Thr Ile Lys Glu Leu Met Arg Trp Tyr 20 25
30 Cys Leu Asn Thr Asn Thr His Gly Cys Arg Arg Ile
Val Val Ser Arg 35 40 45
Gly Arg Leu Arg Arg Leu Leu Trp Ile Gly Phe Thr Leu Thr Ala Val
50 55 60 Ala Leu Ile
Leu Trp Gln Cys Ala Leu Leu Val Phe Ser Phe Tyr Thr 65
70 75 80 Val Ser Val Ser Ile Lys Val
His Phe Arg Lys Leu Asp Phe Pro Ala 85
90 95 Val Thr Ile Cys Asn Ile Asn Pro Tyr Lys Tyr
Ser Thr Val Arg His 100 105
110 Leu Leu Ala Asp Leu Glu Gln Glu Thr Arg Glu Ala Leu Lys Ser
Leu 115 120 125 Tyr
Gly Phe Pro Glu Ser Arg Lys Arg Arg Glu Ala Glu Ser Trp Asn 130
135 140 Ser Val Ser Glu Gly Lys
Gln Pro Arg Phe Ser His Arg Ile Pro Leu 145 150
155 160 Leu Ile Phe Asp Gln Asp Glu Lys Gly Lys Ala
Arg Asp Phe Phe Thr 165 170
175 Gly Arg Lys Arg Lys Val Gly Gly Ser Ile Ile His Lys Ala Ser Asn
180 185 190 Val Met
His Ile Glu Ser Lys Gln Val Val Gly Phe Gln Leu Cys Ser 195
200 205 Asn Asp Thr Ser Asp Cys Ala
Thr Tyr Thr Phe Ser Ser Gly Ile Asn 210 215
220 Ala Ile Gln Glu Trp Tyr Lys Leu His Tyr Met Asn
Ile Met Ala Gln 225 230 235
240 Val Pro Leu Glu Lys Lys Ile Asn Met Ser Tyr Ser Ala Glu Glu Leu
245 250 255 Leu Val Thr
Cys Phe Phe Asp Gly Val Ser Cys Asp Ala Arg Asn Phe 260
265 270 Thr Leu Phe His His Pro Met His
Gly Asn Cys Tyr Thr Phe Asn Asn 275 280
285 Arg Glu Asn Glu Thr Ile Leu Ser Thr Ser Met Gly Gly
Ser Glu Tyr 290 295 300
Gly Leu Gln Val Ile Leu Tyr Ile Asn Glu Glu Glu Tyr Asn Pro Phe 305
310 315 320 Leu Val Ser Ser
Thr Gly Ala Lys Val Ile Ile His Arg Gln Asp Glu 325
330 335 Tyr Pro Ser Val Glu Asp Val Gly Thr
Glu Ile Glu Thr Thr Met Val 340 345
350 Thr Ser Ile Gly Met His Leu Thr Glu Ser Phe Lys Leu Ser
Glu Pro 355 360 365
Ser Ser Gln Cys Thr Glu Gly Gly Ser Asp Val Pro Ile Arg Asn Ile 370
375 380 Tyr Asn Ala Ala Tyr
Ser Leu Gln Ile Cys Leu His Ser Cys Phe Gln 385 390
395 400 Thr Lys Met Val Glu Lys Cys Gly Cys Ala
Gln Tyr Ser Gln Pro Leu 405 410
415 Pro Pro Ala Ala Asn Tyr Cys Asn Tyr Gln Gln His Pro Asn Trp
Met 420 425 430 Tyr
Cys Tyr Tyr Gln Leu His Arg Ala Phe Val Gln Glu Glu Leu Gly 435
440 445 Cys Gln Ser Val Cys Lys
Glu Ala Cys Arg Phe Lys Glu Trp Thr Leu 450 455
460 Thr Thr Ser Leu Ala Gln Trp Pro Ser Val Val
Ser Glu Lys Trp Leu 465 470 475
480 Leu Pro Val Leu Thr Trp Asp Gln Gly Arg Gln Val Asn Lys Lys Leu
485 490 495 Asn Lys
Thr Asp Leu Ala Lys Leu Leu Ile Phe Tyr Lys Asp Leu Asn 500
505 510 Gln Arg Ser Ile Met Glu Ser
Pro Ala Asn Ser Ile Glu Met Leu Leu 515 520
525 Ser Asn Phe Gly Gly Gln Leu Gly Leu Trp Met Ser
Cys Ser Val Val 530 535 540
Cys Val Ile Glu Ile Ile Glu Val Phe Phe Ile Asp Phe Phe Ser Ile 545
550 555 560 Ile Ala Arg
Arg Gln Trp Gln Lys Ala Lys Glu Trp Trp Ala Trp Lys 565
570 575 Gln Ala Pro Pro Cys Pro Glu Ala
Pro Arg Ser Pro Gln Gly Gln Asp 580 585
590 Asn Pro Ala Leu Asp Ile Asp Asp Asp Leu Pro Thr Phe
Asn Ser Ala 595 600 605
Leu His Leu Pro Pro Ala Leu Gly Thr Gln Val Pro Gly Thr Pro Pro 610
615 620 Pro Lys Tyr Asn
Thr Leu Arg Leu Glu Arg Ala Phe Ser Asn Gln Leu 625 630
635 640 Thr Asp Thr Gln Met Leu Asp Glu Leu
645
User Contributions:
Comment about this patent or add new information about this topic:
People who visited this patent also read: | |
Patent application number | Title |
---|---|
20170049643 | MOTOR VEHICLE WITH WHEELCHAIR CADDY |
20170049642 | POWERED COTS |
20170049641 | PROTECTIVE COVER |
20170049640 | PACKAGED ABSORBENT ARTICLE |
20170049639 | UNDERPANTS-TYPE ABSORBENT ARTICLE |